Re: [Histonet] Re: 100% ethanol in tissue processing

[Gudrun Lang via Histonet]

   If there is a cytology lab near you, they will take use of it.
   --
   Diese Nachricht wurde von meinem Android Mobiltelefon mit GMX Mail
   gesendet.

   Am 30.05.24, 01:21 schrieb Curt Tague via Histonet
   :

 My mistake, it’s methanol….
 No bueno for tissue?
 Curt
 Get Outlook for iOS<[1]https://aka.ms/o0ukef>
 
 From: Mac Donald, Jennifer 
 Sent: Wednesday, May 29, 2024 3:50:05 PM
 To: Curt Tague ;
 Histonet@lists.utsouthwestern.edu
 
 Subject: RE: 100% ethanol in tissue processing
 Absolutely can be used on the tissue processor or for staining.
 -Original Message-
 From: Curt Tague via Histonet 
 Sent: Wednesday, May 29, 2024 12:16 PM
 To: Histonet@lists.utsouthwestern.edu
 Subject: [Histonet] 100% ethanol in tissue processing
 EXTERNAL SENDER - Exercise caution with requests, links, and
 attachments.
 Hi all, stupid question (despite what all the college professors
 said, yes, I think there are some stupid questions and this is one
 of them, I should know)!
 Someone dropped off a bunch of pure ethanol, they don't need it, I
 said I'd help dispose it... can't be used for processing can it? I
 seem to recall damaging some tissue many years ago when using
 ethanol... your thoughts?
 Maybe just clean cycles on the processors...
 Thanks,
 Curt
 ___
 Histonet mailing list
 Histonet@lists.utsouthwestern.edu
 [2]https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.utsouth
 western.edu_mailman_listinfo_histonet=DwIFAg=euGZstcaTDllvimEN8b
 7jXrwqOf-v5A_CdpgnVfiiMM=2t7_gY_QgCsQnEhIbW0hGqXohzrIjiJRpXu4w5m7Y
 EQ=xxu_LXkpPKCAe9P_kzXI7q9zsKCFyArunXxppJEo5id6FvQUUNcbma3PfyW68f1
 O=in66vWsozwvAeChBgTJlM8ot_HrKxoikVr2MvJTre_I=
 ___
 Histonet mailing list
 Histonet@lists.utsouthwestern.edu
 [3]http://lists.utsouthwestern.edu/mailman/listinfo/histonet

References

   1. https://aka.ms/o0ukef
   2. 
https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.utsouthwestern.edu_mailman_listinfo_histonet=DwIFAg=euGZstcaTDllvimEN8b7jXrwqOf-v5A_CdpgnVfiiMM=2t7_gY_QgCsQnEhIbW0hGqXohzrIjiJRpXu4w5m7YEQ=xxu_LXkpPKCAe9P_kzXI7q9zsKCFyArunXxppJEo5id6FvQUUNcbma3PfyW68f1O=in66vWsozwvAeChBgTJlM8ot_HrKxoikVr2MvJTre_I=
   3. http://lists.utsouthwestern.edu/mailman/listinfo/histonet
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] [EXTERNAL] Epredia xylene substitute

[Gudrun Lang via Histonet]

Hi Ann,
we have used ShellSol for years in our VIPs until our supplier stopped it. 
That's also a kind of naphta with aliphatic hydrogencarbons.
After that wie changed to the Sakura-substitute with the same protocol. For 
standard processing we had/have two stations with 60 min each at 40 °C. For fat 
processing the time is doubled.
That works for our specimens well enough.
Gudrun

-Ursprüngliche Nachricht-
Von: Ann Specian via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Freitag, 26. April 2024 02:07
An: Will Cavett
Cc: Histonet
Betreff: Re: [Histonet] [EXTERNAL] Epredia xylene substitute

Hi Will, great to hear from you!Yes those are all the reasons why we are 
considering it, but we would like to start validating tissue and rather than 
reinvent the wheel, We are looking for Processing protocols that have already 
been tried and true. If you have any,  if you could share them, it would be 
greatly appreciated.


Sent from the all new AOL app for iOS


On Thursday, April 25, 2024, 6:39 PM, Will Cavett  wrote:

I hope you are doing well, Ann? We considered Epredia's because it seems to 
offer a safer and more convenient option than xylene. Also, the evaporation 
rate of Epredia's Xylene Substitute is comparable to that of xylene, ensuring 
consistent performance in tissue processing .And, unlike xylene, which has a 
strong odor, Epredia's Xylene Substitute is virtually odorless. And lastly, Its 
airborne exposure limit is three times safer than xylene. Hope this helps.

Will Cavett, II

-Original Message-
From: Ann Specian via Histonet 
Sent: Wednesday, April 24, 2024 2:00 PM
To: histonet@lists.utsouthwestern.edu
Subject: [EXTERNAL][Histonet] Epredia xylene substitute

We are thinking of changing from xylene to this substitute. Does anybody have 
any Processing protocols using Epredia Xylene substitute that they could share?


Sent from the all new AOL app for iOS
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Individuals who have received this information in error or are not authorized 
to receive it must promptly return or dispose of the information and notify the 
sender. Those individuals are hereby notified that they are strictly prohibited 
from reviewing, forwarding, printing, copying, distributing or using this 
information in any way.



___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Tissue Processor Down Question

[Gudrun Lang via Histonet]

Depending on the time the cassettes were in clearing medium, I would
transfer them into molten paraffin at least for double of the time in the
processor. And then embed etc.
Try a few blocks to see, if infiltration was sufficient. If cutting is
hampered, prolong the time in paraffin.
Gudrun

-Ursprüngliche Nachricht-
Von: Diana Martinez-Longoria via Histonet
[mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Donnerstag, 14. März 2024 13:59
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Tissue Processor Down Question
Wichtigkeit: Hoch

Good morning,

Our VIP5 tissue processor went down and our tissue cassettes were in
clearing solution. What is the best media to put them while we wait for the
tissue processor to be repaired, please let me know?
Thank you,
Diana Martinez-Longoria
El Centro Regional Medical Center
Lead Histotechnician (ASCP)cm
Laboratory - Pathology Department
1415 Ross Ave | El Centro, CA  92243
760.339.7267: Fax: 760-3394570
 diana.martinez-longo...@ecrmc.org




ECRMC Confidentiality Notice: This e-mail is for the sole use of the
intended recipient(s) and may contain confidential and privileged
information. Any unauthorized review, use, disclosure, or distribution is
prohibited. If you are not the intended recipient, PLEASE contact the sender
and promptly destroy this e-mail and its attachments. 
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] preordering giemsa stains

[Gudrun Lang via Histonet]

In my place it is ordered on demand, but the pathologists usually perfer IHC
for Hp.

Gudrun

-Ursprüngliche Nachricht-
Von: Paula via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Dienstag, 27. Februar 2024 20:46
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] preordering giemsa stains

Hello,

Is it a common practice to preorder stains that involve the stomach
searching for H pylori?  Or, is it more common to order as needed?

Thank you for anyone's feedback.

Paula Lucas

Lab Manager

 

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] microwave processing

[Gudrun Lang via Histonet]

Dear histonetters!

I have a question for those who have an insight in the whole landscape of
pathologies.

I am reading the microwave application book of Dr. Leong (2009). He writes
very enthusiastic about fixation and processing with mircrowaves.

I know there are microwave processors for continous workflow on the market.

 

Now I am curious, how many pathologies use this technology. What do you
think? A few percent or rising numbers?

Thanks in advance and kind regards

Gudrun

 

 

Gudrun Lang

Landgutstraße 25

4040 Linz

 

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] tissue cassettes

[Gudrun Lang via Histonet]

Hi,
Since we have turned to embedding centers in the late 80ies we let the
cassettes sit in the centers without additional paraffin.
We only see such "jumping out" tissue, when the cassettes are not warmed
(let the lid open) and the tissue renders too cold. As a result tissue and
paraffin don't combine well enough.
Maybe it is a matter of embedding technique? Too little paraffin in the mold
before setting the tissue in? Cold embedding molds? Slow handling?
Gudrun

-Ursprüngliche Nachricht-
Von: Brazie, Jeneanne E *HS via Histonet
[mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Freitag, 9. Februar 2024 11:41
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] tissue cassettes

Hello :) I am encountering push back in our lab when I fill the embedding
units with melted paraffin
in the embedding wells. The techs here like for the tissue cassettes  to sit
dry (no wax) while in the
 embedding units. I find that the tissue rolls out of the sections while
cutting because of a layering
effect between the tissue and the paraffin its embedded in. I have
communicated
this but they tell me I'm "old school". Does anyone have any thoughts or
opinions on this topic??

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Bone samples

[Gudrun Lang via Histonet]

Hi,
In my opinion the hardness of the decalcified blocks is often rather due to the 
paraffin-processing than the residual calcium. Especially when the tissue is 
decalcified really long. The hardness comes from the dehydration and "cooking" 
of collagen fibers. So additional decal will not help reducing the calcium, but 
helps to reintroduce water into the collagen-grid. And this is helpful for 
softening and cutting.
For myself, I often scratch the paraffin on the blocksurface away to face the 
bone directly to the water. Then I let them "swim" on my waterbath, until the 
surface is turned rather milky. After cooling again I cut in very very small 
steps to trim the surface. Sometimes it needs repeated swimming and cooling 
(and patience) to get a rather acceptable section. It is advantageous to pick 
them up on adhesive slides and let them dry in an 60°C oven to get rid of any 
residual water under the section.

Hope this helps
Gudrun

-Ursprüngliche Nachricht-
Von: Chakib Boussahmain via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Mittwoch, 24. Jänner 2024 23:34
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Bone samples


Hi guys,

 I hope this messagefinds you well. I am currently working on a study involving 
bone samples thathave been treated with slow decal and embedded in paraffin. I 
am facingchallenges in obtaining nice sections, and I was wondering if you 
could providesome guidance or recommendations.

Thank you in advance for your help!
Chakib
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Detergent in heating antigen retrieval

[Gudrun Lang via Histonet]

Hi,
I think the answer is  as so often: it depends. Detergens solves membranes 
partly and leads to a higher permeabilization of the tissue. Some antigens may 
take advantage of that, some may not need it. 
Higher permeability is good for detection with high molecular complexes. 

Gudrun Lang

-Ursprüngliche Nachricht-
Von: Alonso Martínez Canabal via Histonet 
[mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Mittwoch, 20. September 2023 21:25
An: Histonet
Betreff: [Histonet] Detergent in heating antigen retrieval

Dear histoneters,
   I have performed heating antigen retrieval with citrate buffer
pH 6 with 0.05% tween-20, however I have seem recipes  with no detergent,
anyone has any experience or knowledge if it is better with or without the
detergent?
  Thank you!

-- 
Dr. Alonso Martínez Canabal PhD
Profesor Asociado "C"
Departamento de Biología Celular, Facultad de Ciencias, UNAM
Investigador Nacional "I"
56224833
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Prostate biopsy-water logged popping /lost at cutting?

[Gudrun Lang via Histonet]

What first comes in my mind is, that the tissue was a little too cold when
placed into the embedding mold. That would cause an insufficient junction
with the paraffin.
If there was not enough hot paraffin in the embedding mold and handling took
a little bit too long, or if the tissue was rather cold and dry before
putting it into the mold (cassette too long open before embedding)
something like that.
Too brittle tissue because of the processing or because it was airdried
before putting it into formalin, tends to pop out of the block. But this
would lead to obvious cutting problems.

Regards
Gudrun

-Ursprüngliche Nachricht-
Von: Mona Amin via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Donnerstag, 20. Juli 2023 23:36
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Prostate biopsy-water logged popping /lost at cutting?

Hi everyone,

We had a case of prostate core biopsies total 12 blocks

First 2 block cut fine and when cutter touched third block lost the core
completely. The blocks were on ice face down no longer than 20 mins once
faced.
The other blocks looked like they were popping out and were taken for
re-embedding and then sectioned.

Thoughts if this popping out could be because of
1- too much time in water
2- processing issue
3-condesnsation issue from cold plate

Generally too we see popping of our biopsies but with this case it was
worst.
Any help/experiences shared will be greatly appreciated as we try to
resolve the root cause.

Thank you everyone in advance.
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] shelf life of working antibody solutions in IHC

[Gudrun Lang via Histonet]

Hi all!

I have tried to find a general instruction for the shelf life of antibody
working solutions.

With automated IHC you usually fill the container with the working solution
and depending on the frequency of usage they stay on the instrument at
roomtemperature (or higher) or are put in the fridge again.

The working solutions are up to 10 ml and may last for months. The
antibody-diluent is from the same company of the instrument. The titers are
in a range from 1:10 to 1:3000. - so a very heterogen situation.



How do you handle this? Have you a general rule, when the solution has to be
discarded? Is it just a matter of positiv-controls?



Thanks in advance

Gudrun Lang

Biomedical scientist

Austria





___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] history of H staining

[Gudrun Lang via Histonet]

Hi all!

Does anybody know, when the H stain became that dominant routine-stain in
the pathology labs?

It was introduced by Wissowzky 1876, but I am curious when our usual
histoprocess became worldwide standard.



Regards

Gudrun Lang

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] cutting with glassknifes

[Gudrun Lang via Histonet]

Dear histonetters!

I have a question for the experienced histo-people.

Is it still in practice to use glassknifes for cutting on rotary- or
sliding-microtomes? For plastic-embedded tissue in light-microscopy?

Or ar glassknifes only found in EM-technique?



I am just curious, because I assume, it is a rather difficult skill to
handle.

I made a short internet-search, but got no hints on this issue.



Thank you and kind regards

Gudrun Lang

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Sudanblack B on FFPET

[Gudrun Lang via Histonet]

Many thanks to all for your helpful hints.

Kind regards
Gudrun Lang

-Ursprüngliche Nachricht-
Von: Tony Henwood via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Dienstag, 28. März 2023 21:13
An: Bob Richmond; Histonet@lists.utsouthwestern.edu
Betreff: Re: [Histonet] Sudanblack B on FFPET

I would also let the saturated solution stand for a few days. Like Oil Red
O, it probably needs time to “mature”. I would also use a frozen section of
skin as a control.

Regards,

Tony Henwood
Sydney, Australia

From: Bob Richmond via Histonet<mailto:histonet@lists.utsouthwestern.edu>
Sent: Wednesday, 29 March 2023 4:51 AM
To:
Histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>
Subject: Re: [Histonet] Sudanblack B on FFPET

>
> Gudrun Lang in Austria asks:
>

>>Has anyone experience with Sudanblack B on paraffin slides for staining
[lipofuscin]? A doctor wants the demonstration of the lipoid content of
foamy cells or granulocytes in lung. I've found protocols that have
incubation-times from 10 minutes to over-night. - I've made a saturated
Sudan black B-solution in 70% ethanol and tried it with10 min on liver
without real success.<<

The main thing you need to do is demonstrate that it isn't hemosiderin with
an iron stain (Perls prussian blue reaction), and perhaps also that it
isn't acid-fast. Lipofuscin can be identified an H & E staining, except for
these considerations.

Bob Richmond
Maryville, Tennessee
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Sudanblack B on FFPET

[Gudrun Lang via Histonet]

Hallo!

Has anyone experience with Sudanblack B on paraffin slides for staining
lipofuszin? A doctor wants the demonstration of the lipoid content of foamy
cells or granulocytes in lung.



I've found protocols that have incubation-times from 10 minutes to
over-night.

I've made a saturated Sudanblack B-solution in 70% ethanol and tried it with
10 min on liver without real success.



I would appreciate any input and help.

Thanks in advance



Gudrun Lang

Austria

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] destain for IHC

[Gudrun Lang via Histonet]

We do restaining for IHC also, but without destaining at all. Hematoxylin
doesn't matter because nuclei are stained after IHC with hematoxylin anyway.
And Eosin gets lost through all the washing in buffer.
The problem is, that HE section are usually mounted on non-adhesive slides,
and they float away during the procedure.
Gudrun

-Ursprüngliche Nachricht-
Von: Nancy Schmitt via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Mittwoch, 8. Februar 2023 18:29
An: Histonet
Betreff: [Histonet] destain for IHC

Hello-

Has anyone had success destaining an H and then running IHC on the same
slide?

Thanks!

Nancy


Confidentiality Notice:
This e-mail, including any attachments is the property of Trinity Health and
is intended for the sole use of the intended recipient(s). It may contain
information that is privileged and confidential.  Any unauthorized review,
use, disclosure, or distribution is prohibited. If you are not the intended
recipient, please delete this message, and reply to the sender regarding the
error in a separate email.
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Alcianblue for fast frozen sections

[Gudrun Lang via Histonet]

Dear histonetters!

Today I've heard about alcianblue staining on fast frozen sections, but I've
got no details.

I would like to know, if the staining result is the same as for staining AB
on paraffinslides.



They use the stain on transplantation liver.

Is this a usual procedure?



I would be glad about any information.

Thanks in advance

Gudrun

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Reconstituting antibodies and dilutions

[Gudrun Lang via Histonet]

500 µg in 1000µl  is the same as 100µg in 200µl

So I would reconstitute the 100 µg with 200 µl of water to get the liquid
antibody-concentrate.
Then I would make the working-dilution: eg. 5µl of concentrate + 5000µl
buffer 

The correct way would be 5 µl + 4995 µl, but in practice this does'nt
matter.

Gudrun Lang

-Ursprüngliche Nachricht-
Von: Charles Riley via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Donnerstag, 13. Oktober 2022 17:49
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Reconstituting antibodies and dilutions

When calculating the dilution of an antibody when do start?

i.e.  If i have 100 ug of lyophilized antibody and its recommended
reconstitution is 500 ug/ml

How do I mix a 1:1000 dilution?

I am thinking it's adding 2mL diH20 to create the 500 ug/mL and
then diluting the sample.
And I would be able to make 2000ml of 1:1000 concentration solution ?  2mL
antibody and 1998mL diluent?


Please let me know if I made a calculation error
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Absence of Crystals in stains

[Gudrun Lang via Histonet]

My first thought about calciumcrystals is solving by the chromatic acid.
I would recommend a vonKossa stain  for calcium deposits.

Regards Gudrun Lang

-Ursprüngliche Nachricht-
Von: Akemi via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Dienstag, 9. August 2022 23:51
An: Histonet
Betreff: [Histonet] Absence of Crystals in stains

I received this from a pathologist friend and need your help.  Unfortunately, I 
can’t attach the images he sent me on histonet.
“I’ve got a histopuzzle for you. This is a fungus ball from a maxillary sinus. 
On the H, it is loaded with these birefringent crystals, which I hypothesize 
are calcium oxalate. On the GMS stain, however, the crystals are completely 
absent. 

Do you think there’s some reagent in the GMS procedure that dissolves crystals 
that would be resistant to tissue processing and the H stain? “

Thanks in advance!
Akemi Allison BS, HT/HTL

Sent from my iPhone
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Looking for old Microm-bladeholders

[Gudrun Lang via Histonet]

Dear histonetters,

I am looking for old Microm bladeholders for sliding-microtomes. Those ones,
that have a shorter coverplate than the standard blade. We prefer them for
working, but they aren't sold any more.

I would be happy for any idea or contact - preferable in Europe. Maybe
someone has an unused bladeholder in the lab-storage.



Thanks in advance



Lang Gudrun

Austria

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Prussian Blue Reaction

[Gudrun Lang via Histonet]

Hi Jennifer,
Why do you want to reduce the staining?

I ask, because the impact of hydrochloric acid on the tissue may influence
the following results anyway.
I think, that the prussian blue pigment cannot be removed in an easy way. It
is resistent to solvents and mineral acids.
http://www.epsilonpigments.com/inorganic-pigment/prussian-blue/Prussian-Blue
-for-Solvent-Based-Inks.html

On the other hand, if the blue colour doesn't interfere with your following
staining, you can try to simple make a "double stain".

Regards
Gudrun

-Ursprüngliche Nachricht-
Von: Mac Donald, Jennifer via Histonet
[mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Sonntag, 6. Juni 2021 06:34
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Prussian Blue Reaction


Does anyone know of a way to remove/reduce the Prussian blue reaction?
Thanks,
Jennifer



___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] hexamethyleneimine

[Gudrun Lang via Histonet]

Jones Methenamin Silver Impregnation, for staining basalmembrans in
glomeruli.
Gudrun

-Ursprüngliche Nachricht-
Von: LEROY H BROWN via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Montag, 12. Oktober 2020 00:45
An: histonet@lists.utsouthwestern.edu
Betreff: Re: [Histonet] hexamethyleneimine

What do you use hexamethyleneimine for in your lab.   I found an old bottle
of this reagent and cannot recall why I have it.

It must be used in making up a stain but I am not remembering what stain.

Anyone know?

Thanks

LeRoy Brown HT(ASCP)HTL

 

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Spectra

[Gudrun Lang via Histonet]

Hi!
Do you use your own reagens or the Leica Spectra dyes? We have house-made
ones.
Too faint eosin staining can be rendered more intense with a few drops of
acetic acid into the eosin to reach a pH about 5. Second, it is often a
prolonged washing after the eosin that causes excess stain to be lost.

Our protocol is 1 min in 2% aequoues eosin (pH 5,5), rinsing in running
tapwater for 1 min, then 1 min in 96% ethanol,  2x 100%, xylen. The times
for rinsing in water and 96% have to be "exact" in the protocol. In 100% and
ethanol a prolonged stay doesn't matter.
We optimized the turnaraound time with two identical protocols for HE, that
use two staining dishes each for hematoxylin and eosin. The rest of the
dishes is used by both protocols. Except of the former mentioned stations,
the rest is "not exact". So the Spectra has more "ways" to find an optimal
protocol.
The all-over staining time is also about 60 min, due to 25 min in the oven
and the progressive hematoxylin (10 min). Usually we feed the stainer
continuosly  in 15-20 min steps, as the racks are filled after cutting. And
the racks are ready in the same pattern.

Here, we have no problems with too fast lowering of the reagens-level.

Hope, this helps.
Gudrun

-Ursprüngliche Nachricht-
Von: warda hassan via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Sonntag, 19. April 2020 11:37
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Spectra

Good Morning to all

Thanks in advance to all who would help with their feedback on the
automation of H/E staining,
Initially we have installed Spectra-Leica for H/E
The feedback from pathologist is the stain is bit on bluish and eosin is
weak
We did altered and reached to 2 min of Hematoxylin and 11 min Eosin
But unfortunately the stain consistency is not stable as after every 2-3
racks level of reagents reduces so we top up as we do the intensity of it
differs
Second issue is Turn around time, each rack takes 45mins, so tried adding
up new protocol as copy but the machine keeps a gap of 12mins in picking up
the slides for staining !
If anyone of you have experienced with Leica automation
Please your feedback on above would be of a great help
Thank you so much
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Question concerning H. pylori staining

[Gudrun Lang via Histonet]

Hi!
I found this instruction for a Hp- stain, that sounds similiar to yours.
They want the slides to be airdried after water-rinsing and before xylen.
But the result should be blue bacteria, not purple.
I would try to let the slides air-dry for about half a minute, then rinse
very-very short in absolute ethanol (one dip). The colour should turn blue
and some dye will be extracted to give a clearer result. Then fast into
xylen, dip a few times, then coverslip.

http://www.helicostat.com/helicostat_instructions.html
I assume, that the periodic acid should render some mucins stainable with
Alcian yellow, to give a more contrasted result.

Gudrun Lang



-Ursprüngliche Nachricht-
Von: Bob Richmond via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Donnerstag, 9. April 2020 20:05
An: Histonet@lists.utsouthwestern.edu
Betreff: Re: [Histonet] Question concerning H. pylori staining

Michelle (where) asksi: >>We have a question about staining for H-Pylori
Using Quick Stain (Periodic acid 1%, Alician Yellow, Sodium Metabisulfate,
Toluidine Blue stock, Sodium Hydroxide) we notice what clearly  looks like
the H-Pylori purple stained clusters, but after dehydration in 100% alcohol
the purple clusters seem to disappear. Should we just dehydrate using a
slide oven instead of the alcohol? For how long at what temp? Could the
alcohol be affecting the purple color making it too light to see?<<

You identify Helicobacter pylori by its morphology - curved, angled, or
"gull wing" bacilli. Is that what you're seeing? If you do this stain, you
should know how it's interpreted.

The Alcian yellow (correct spelling) method is needlessly complex. What
does the periodic acid do? - A solution of toluidine blue (Diff-Quik 2 or a
generic equivalent) suffices - don't use Diff-Quik 1 with it - dehydrate
through alcohols rapidly.

Bob Richmond
Samurai Pathologist
Maryville TN
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] FISH on BondMax

[Gudrun Lang via Histonet]

Dear all!

I would appreciate input on the FISH-function of the BondMax from Leica. Is
this feature reliable and better/as good as manual staining?

Would anyone recommend this instrument for FISH staining?



Thank you



Lang Gudrun



___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Stainer vs. Stainer

[Gudrun Lang via Histonet]

Hi Terri,
Have you actually used the Spectra from Leica? This is the follower of
ST5020 and CV5020.
I am also interested in some feedback on this instrument. 

Gudrun Lang

-Ursprüngliche Nachricht-
Von: Terri Braud via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Donnerstag, 14. März 2019 20:10
An: 'histonet@lists.utsouthwestern.edu'
Betreff: [Histonet] Stainer vs. Stainer

Hi Alison - 
I've used both stainers and like both of them a lot.  Both were super
reliable and easy to use.  However, coverslipping is a different story.
I've used both film and glass.  About film - super quick, super easy, but -
the purity of the xylene used to coverslip from film must be absolute.
Anyone who has experienced film pulling off the slides in storage had a
miniscule portion of water carried down the acohols and into the xylene. If
it were glass, the process is a bit more forgiving of water contaminent. The
absolute alcohols leading to the end xylenes must be kept very fresh.  I
kept film slides for over 20 years, no problem.  If you are looking into
digital pathology, I would check with vendors to see if film is acceptable.
I don't know.
As to coverslippers, we've been using the Sakura glass now for 10 years and
love it.  I can't compare it to the newer Leica Glass, but 10 years ago my
techs all preferred the Sakura because it had fewer moving parts and the
maintenance was easier.  I hope this helps.  Good Luck, Terri

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Laboratory
Holy Redeemer Hospital
1648 Huntingdon Pike
Meadowbrook, PA 19046
ph: 215-938-3689
fax: 215-938-3874
Care, Comfort, and Heal

   6. stainer v. stainer (Perl , Alison)
   
Message: 6
Date: Wed, 13 Mar 2019 20:08:14 +
From: "Perl , Alison" 
To: "'histonet@lists.utsouthwestern.edu'"

Subject: [Histonet] stainer v. stainer


Hi all
We are getting ready to purchase a new H stainer/coverslipper, and are
considering the Sakura Prisma Plus (tape) and the Leica Spectra (glass).
Does anyone have good or bad feedback on either instrument, and/or tape v.
glass? We've always had glass, but of course the coverslippers need more
maintenance, take longer to dry, more expensive than tape, etc etc. So we
are very interested in tape, but still a little hesitant about the old
problems of yellowing and peeling after 10+ years. Since we're in NY, we
have to keep all slides for 20 years

Any thoughts are appreciated!

Alison Perl, HTL(ASCP)CM
Anatomic Pathology Manager
CareMount Medical
110 South Bedford Rd
Mount Kisco, NY 10549
(914) 302-8424
ap...@cmmedical.com<mailto:ap...@cmmedical.com>
www.caremountmedical.com<http://www.caremountmedical.com/>



___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] FISH question

[Gudrun Lang via Histonet]

Thanks for your advice.
I use adhesive slides from Leica. Years ago we had Superfrosts of another
brand (maybe Thermo) and I can't remember similar issues.
Can you recommend a special brand for FISH?
Have you ever tried to do FISH on a slide without adhesion?

Gudrun

-Ursprüngliche Nachricht-
Von: Whitaker, Bonnie [mailto:bonnie.whita...@osumc.edu] 
Gesendet: Dienstag, 19. Februar 2019 18:54
An: 'Mark Tarango'; Gudrun Lang
Betreff: RE: [Histonet] FISH question

I know a few years ago, we ran into the same issue, and the problem actually
was with the slides.  We were using cheaper slides for most of histology at
the time, but had to purchase "higher end" slides for the FISH.

Thanks,
Bonnie

Bonnie P. Whitaker
AP Operations Director

The Ohio State University
Wexner Medical Center
Department of Pathology
N305 Doan Hall
410 West 10th Avenue
Columbus, Ohio  43210

614.293.8418
FAX 614.293.2779
Pager: 614.293.7243 ext. 5013

-Original Message-
From: Mark Tarango via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Tuesday, February 19, 2019 12:43 PM
To: Gudrun Lang
Cc: HistoNet
Subject: Re: [Histonet] FISH question

Hi Gudrun,

Are you sure you have digested long enough with pepsin?  If the tissue is
not well digested you will see background.  We use sodium thiocyanate for
pretreatment reagent, not citric buffer.  These are my first thoughts.

Mark

On Tue, Feb 19, 2019 at 9:18 AM Gudrun Lang via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Dear histonetters!
>
> I have difficulties with my FISH preparation on FFPET. I struggle with
> massive background. It looks like a thick fluorescent film.
>
> The signals can't be seen because of the background. Even the nuclei are
> hard to see.
>
> The background is within the tissue but also surrounds it. Therefore it
> must
> be directly on the glass slide.
>
>
>
> The slide is clear after deparaffination and after pretreatment with
citric
> buffer and pepsin. After the pepsin the slides are rinsed in 2xSSC, then
> 50%-70%-96%-100% ethanol (p.a.).
>
> And then the slides are airdried.
>
> On the dry slides foggy streams appear. The slides become turbid. When I
> rinse them again in graded ethanols it becomes better but still a little
> turbid.
>
> After hybridisation and stringent washing the slides are air-dried again
> and
> coverslipped with Dapi.
>
> When looking at the slides in the fluorescence microscope the trouble
> arises.
>
>
>
> My assumption is, that there is a remnant of the salt of the SSC buffer.
> How
> can I inhibit this deposit? Can I replace the buffer with water without
any
> harm to the tissue?
>
> Or ist there a different cause for the turbidiy?
>
> I use fresh reagenses from xylene to buffer and ethanol.
>
> Any hints are welcome.
>
>
>
> Thanks in advance
>
> Gudrun Lang
>
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
>
https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.utsouthwestern.edu
_mailman_listinfo_histonet=DwICAg=k9MF1d71ITtkuJx-PdWme51dKbmfPEvxwt8SFE
kBfs4=BgjUe6oLZB0OAcW6Y6Rn-n0Q03Ac4dWC2x8Sg24AeiY=gzHjMQcM0W22Saad2E8Pgv
a_UOfvxrD1xD0IQ57lDUY=rVkT1YjvFe8uXhvIGPusCI6h-qfQm2I-v86i1XIRePc=
>
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.utsouthwestern.edu
_mailman_listinfo_histonet=DwICAg=k9MF1d71ITtkuJx-PdWme51dKbmfPEvxwt8SFE
kBfs4=BgjUe6oLZB0OAcW6Y6Rn-n0Q03Ac4dWC2x8Sg24AeiY=gzHjMQcM0W22Saad2E8Pgv
a_UOfvxrD1xD0IQ57lDUY=rVkT1YjvFe8uXhvIGPusCI6h-qfQm2I-v86i1XIRePc=


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] FISH question

[Gudrun Lang via Histonet]

Dear histonetters!

I have difficulties with my FISH preparation on FFPET. I struggle with
massive background. It looks like a thick fluorescent film.

The signals can't be seen because of the background. Even the nuclei are
hard to see.

The background is within the tissue but also surrounds it. Therefore it must
be directly on the glass slide.

 

The slide is clear after deparaffination and after pretreatment with citric
buffer and pepsin. After the pepsin the slides are rinsed in 2xSSC, then
50%-70%-96%-100% ethanol (p.a.).

And then the slides are airdried. 

On the dry slides foggy streams appear. The slides become turbid. When I
rinse them again in graded ethanols it becomes better but still a little
turbid.

After hybridisation and stringent washing the slides are air-dried again and
coverslipped with Dapi. 

When looking at the slides in the fluorescence microscope the trouble
arises. 

 

My assumption is, that there is a remnant of the salt of the SSC buffer. How
can I inhibit this deposit? Can I replace the buffer with water without any
harm to the tissue?

Or ist there a different cause for the turbidiy? 

I use fresh reagenses from xylene to buffer and ethanol. 

Any hints are welcome.

 

Thanks in advance 

Gudrun Lang

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] VIP issue

[Gudrun Lang via Histonet]

Thank you for all the good hints and advices.

It has to be a reagens-issue, because the second instrument was also
concerned. 
But we had no similar problems for decades, although we use 96% as
cleaning-ethanol after ShellSol. 
We change the reagenses after 5 runs, once a week. 

We soon are forced to end using ShellSol, because the company stops the
production. We don't like to switch to xylene. 
Which xylen-substitutes are recommended for the VIP besides of Tissue-Clear
(Sakura)?

Best wishes
Gudrun 


-Ursprüngliche Nachricht-
Von: Gudrun Lang via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Donnerstag, 11. Oktober 2018 18:58
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] VIP issue

Dear all!

I have a question for those, who are familiar with the VIPs from Sakura. 

Last time we changed the reagenses the cleaning-ethanol (96%) was very milky
and even was full of many small particles. It was a paraffin-soup.

What is the cause for such a case?  We have been using the organic solvent
(ShellSol) for decades as xylensubstitute. 

Maybe the quality has suffered and the ability of solving paraffin has
decreased. But are there other explanations? Maybe a malfunction of the
instrument?

 

Thanks in advance

Gudrun Lang

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] VIP issue

[Gudrun Lang via Histonet]

Dear all!

I have a question for those, who are familiar with the VIPs from Sakura. 

Last time we changed the reagenses the cleaning-ethanol (96%) was very milky
and even was full of many small particles. It was a paraffin-soup.

What is the cause for such a case?  We have been using the organic solvent
(ShellSol) for decades as xylensubstitute. 

Maybe the quality has suffered and the ability of solving paraffin has
decreased. But are there other explanations? Maybe a malfunction of the
instrument?

 

Thanks in advance

Gudrun Lang

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Quality Measure for Pathology

[Gudrun Lang via Histonet]

Linda,
that's exactly what we do here.
I wondered, if you fixed a cut-off. Because in quality-audits they often
asked the "what do you if.."-questions. 
for example: "There is an increase in the number of discrepancies in this
special month. The rate is over your defined aim. What did you do to fix the
problem? Do you keep records about the process of solving the problem?"

I was just curious, if you go further than the counting of events. 
In daily work we often know the causes of mistakes, and very often we know,
that a certain amount of mistakes can't be avoided, although everyone tries
to give his/her best. 
I think QM should help to discover the special events or circumstances, that
raise the number of mistakes above the "normal" level. I was curious, if you
defined this "normal" level for your lab.

Kind regards
Gudrun


-Ursprüngliche Nachricht-
Von: Blazek, Linda [mailto:lbla...@digestivespecialists.com] 
Gesendet: Mittwoch, 13. Juni 2018 14:38
An: Gudrun Lang
Cc: histonet@lists.utsouthwestern.edu
Betreff: RE: [Histonet] Quality Measure for Pathology

Gudrun,
Our lab is 95% biopsy specimens.  Therefor the number of pieces counted at
gross is written on the side of the cassette so that at the time of
embedding the tech knows what should be present.  That helps in being sure
there isn't a piece stuck to the lid or forceps, or popped out when opening
the cassette.  It's really more of a Quality Assurance measure than tracking
a discrepancy.  I do watch how frequent there is a discrepancy though and if
there seems to be a pattern will initiate a review of processes for both the
grossing person and the embedding person.  
What we see most is the gross will say one piece and there will actually be
multiple tiny pieces or two pieces will be three at embedding.  As all staff
is aware that we are following this they tend to be much more precise in
counting at grossing and embedding.  
Linda

-Original Message-
From: Gudrun Lang [mailto:gu.l...@gmx.at] 
Sent: Wednesday, June 13, 2018 4:19 AM
To: Blazek, Linda
Cc: histonet@lists.utsouthwestern.edu
Subject: AW: [Histonet] Quality Measure for Pathology

Linda,
How do you handle this key figure of discrepancies? Have you set a cut-off
(per person, per day) for any consequencies? What are the
consequencies, besides repeated training?

thanks
Gudrun

-Ursprüngliche Nachricht-
Von: Blazek, Linda via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Dienstag, 12. Juni 2018 18:55
An: Normington Lacy; Amy Self
Cc: histonet@lists.utsouthwestern.edu
Betreff: Re: [Histonet] Quality Measure for Pathology

Also a discrepancy between the number of pieces that are dictated at gross
as being in a cassette compared to what is found in the
cassette at embedding.  You can record the number of blocks that need
re-embedded and the number of slide recuts needed a month. 

Linda

Linda Blazek HT (ASCP)
Pathology Lab Manager
GI Pathology of Dayton
Digestive Specialists, Inc
Phone: (937) 396-2623
Email: lbla...@digestivespecialists.com

-Original Message-
From: Normington Lacy via Histonet
[mailto:histonet@lists.utsouthwestern.edu] 
Sent: Tuesday, June 12, 2018 9:34 AM
To: Amy Self; 'histonet@lists.utsouthwestern.edu'
Subject: Re: [Histonet] Quality Measure for Pathology

Some ideas:

Proficiency Testing
Corrected Results (Addendums/Amendment) volume
Turnaround Time Monitoring
Frozen Section TAT (critical call)
Rejected Tests (empty bottles, etc)


-Original Message-
From: Amy Self via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Tuesday, June 12, 2018 8:18 AM
To: 'histonet@lists.utsouthwestern.edu' 
Subject: [Histonet] Quality Measure for Pathology

WARNING: This email appears to have originated outside of the UW Health
email system.
DO NOT CLICK on links or attachments unless you recognize the sender and
know the content is safe.




Good Morning,

I am looking for ideas/suggestions for QM Histology/Pathology.  My QM
director wants me to "measure" something that I can place on
the lab dashboard.

Thanks in advance for your help..

Amy Self
Histology Lab Senior Tech
Lab
Tidelands Georgetown Memorial Hospital
606 Black River Road
Georgetown, SC 29440
843-520-8711
as...@tidelandshealth.org
Our mission:  We help people live better lives through better health.
NOTE:
 The information contained in this message may be privileged, confidential
and protected from disclosure. If the reader of this
message is not the intended recipient, or an employee or agent responsible
for delivering this message to the intended recipient,
you are hereby notified that any dissemination, distribution or copying of
this communication is strictly prohibited. If you have
received this communication in error, please notify us immediately by
replying to this message and deleting it from your computer.
Thank you.
___
Histonet mailing list
Histonet@lists.

Re: [Histonet] Quality Measure for Pathology

[Gudrun Lang via Histonet]

Linda,
How do you handle this key figure of discrepancies? Have you set a cut-off (per 
person, per day) for any consequencies? What are the
consequencies, besides repeated training?

thanks
Gudrun

-Ursprüngliche Nachricht-
Von: Blazek, Linda via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Dienstag, 12. Juni 2018 18:55
An: Normington Lacy; Amy Self
Cc: histonet@lists.utsouthwestern.edu
Betreff: Re: [Histonet] Quality Measure for Pathology

Also a discrepancy between the number of pieces that are dictated at gross as 
being in a cassette compared to what is found in the
cassette at embedding.  You can record the number of blocks that need 
re-embedded and the number of slide recuts needed a month. 

Linda

Linda Blazek HT (ASCP)
Pathology Lab Manager
GI Pathology of Dayton
Digestive Specialists, Inc
Phone: (937) 396-2623
Email: lbla...@digestivespecialists.com

-Original Message-
From: Normington Lacy via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Tuesday, June 12, 2018 9:34 AM
To: Amy Self; 'histonet@lists.utsouthwestern.edu'
Subject: Re: [Histonet] Quality Measure for Pathology

Some ideas:

Proficiency Testing
Corrected Results (Addendums/Amendment) volume
Turnaround Time Monitoring
Frozen Section TAT (critical call)
Rejected Tests (empty bottles, etc)


-Original Message-
From: Amy Self via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Tuesday, June 12, 2018 8:18 AM
To: 'histonet@lists.utsouthwestern.edu' 
Subject: [Histonet] Quality Measure for Pathology

WARNING: This email appears to have originated outside of the UW Health email 
system.
DO NOT CLICK on links or attachments unless you recognize the sender and know 
the content is safe.




Good Morning,

I am looking for ideas/suggestions for QM Histology/Pathology.  My QM director 
wants me to "measure" something that I can place on
the lab dashboard.

Thanks in advance for your help..

Amy Self
Histology Lab Senior Tech
Lab
Tidelands Georgetown Memorial Hospital
606 Black River Road
Georgetown, SC 29440
843-520-8711
as...@tidelandshealth.org
Our mission:  We help people live better lives through better health.
NOTE:
 The information contained in this message may be privileged, confidential and 
protected from disclosure. If the reader of this
message is not the intended recipient, or an employee or agent responsible for 
delivering this message to the intended recipient,
you are hereby notified that any dissemination, distribution or copying of this 
communication is strictly prohibited. If you have
received this communication in error, please notify us immediately by replying 
to this message and deleting it from your computer.
Thank you.
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] inhouse procedures in histotechnics

[Gudrun Lang via Histonet]

Hi!

What would you say is an inhouse-procedure in a typical clinical histo-lab. The 
question is regarded to accredidation in quality
management.

 

I'm asked to make a list of all inhouse-procedures. Special staining protocols? 
Immunhisto on stainer? 

I think histolabs are hardly comparable to typical medical/chemical labs with 
mainly commercial kits on instruments. 
Changing some duration of a staining incubation to get a better result - is 
this inhouse-made then??

What is the golden standard of all our stainings to refer to? Giemsa 1901? 
Masson 1929? Klebs (Paraffin embedding) 1871?
Wissowzky,Busch (H) 1876?   Mayer (hemalaun) 1891?  ... and so on; you know 
what I mean?

 

I think this is similar to a restaurant-kitchen, where each recipe is usually 
inhouse-made, although everyone is cooking "Pasta". 
Especially if you do the stainings by hand. (grumbling)

 

I know QM makes the lab-world better, but sometimes it is too much for me. ; )

 

Would be happy to get some input.

thanks in advance

 

Gudrun Lang

Austria

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Bouin postfixation

[Gudrun Lang via Histonet]

The treatment with bouin afterwards doesn't make the formaldehyde-fixation 
undone. This would be rather a kind of antigen-retrieval
and results at least in a mixture of both fixations. 
I would stay with the familiar formaldehyde-fixation without experiments and 
try to get the best out of it with the best fitting
antigen-retrieval method.

Gudrun Lang

-Ursprüngliche Nachricht-
Von: SANCHEZ QUINTEIRO PABLO via Histonet 
[mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Sonntag, 3. Juni 2018 20:01
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Bouin postfixation


Dear histonetters,

I have to do immunohistochemistry with an antibody that works better in Bouin 
fixed samples. But I am afraid that my samples -just
unprocessed- are in formalin.

Do you think that it could be help to transfer them to Bouin 24 hours and then 
70 ethanol and embedding?

Thanks in advance

Pablo Sanchez-Quinteiro
Lugo. Spain
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] RET IHC testing

[Gudrun Lang via Histonet]

I bought it from Cellsignalling (monoclonal rabbit), but just started with the 
procedure on lung tissue with no experience until
now.
I took ileum as positiv control, with stained Peyer-plaques as result.

Gudrun Lang

-Ursprüngliche Nachricht-
Von: Charles Riley via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Freitag, 1. Juni 2018 21:07
An: Histo List
Betreff: [Histonet] RET IHC testing

Does anyone perform this testing?

If so who do you get your antibody from and what platform do you run it on.
Trying to figure out where to buy the antibody from. Looking for thoughts
on cost and ease of use from suppliers.

I use the Leica Bond platforms

-- 

Charles Riley BS  HT, HTL(ASCP)CM

Histopathology Coordinator/ Mohs
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] V600E

[Gudrun Lang via Histonet]

This is an mutationspecific antibody against BRAF. And the mutation detected is 
called p.V600E. The exchange of an nucleotid in the
DNA (codon 600) leads to an tranformed and always activated BRAF-protein, that 
is sensitive to an thyrosinkinase-inhibitor as an eg.
Melanoma therapy. As far as I remember it was sold by Novus Biological before 
Roche-Ventana had it. The clone is VE1.

Gudrun

-Ursprüngliche Nachricht-
Von: Normington Lacy via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Mittwoch, 6. Juni 2018 16:43
An: Laurie Colbert; 'histonet@lists.utsouthwestern.edu'
Cc: jtouchst...@pathmdlabs.com
Betreff: Re: [Histonet] V600E

I am thinking this might be BRAF, clone V600E.  We get ours from Ventana Roche.

Lacy Normington
UW Health 
Madison, WI


-Original Message-
From: Laurie Colbert via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Wednesday, June 06, 2018 8:50 AM
To: histonet@lists.utsouthwestern.edu
Cc: jtouchst...@pathmdlabs.com
Subject: [Histonet] V600E

WARNING: This email appears to have originated outside of the UW Health email 
system.
DO NOT CLICK on links or attachments unless you recognize the sender and know 
the content is safe.




Can anyone tell me about the V600E antibody?  Our pathologist wants to add it 
to our menu of IHC stains, and I've never heard of it.
Sources?

Thanks,
Laurie Redmond
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Masson's Trichrome Troubleshooting

[Gudrun Lang via Histonet]

Hi,
Check the staining after the PTA/PMA step. The acid should destain all the
tissue areas, where afterwards the anilinblue should bind. The acid replaces
the Biebrich scarlett in this areas and enables and enhances the binding of
anilinblue.
If the differentiation in insufficient, it may be the quality of the acid to
be the culprit. The pH should be about 2, if I remember correctly.

Check the temperature of the bouin after the microwave. If you cook the
tissue, it may also change the stainability. Microwave is not microwave, the
instrument in your old lab may have different characteristics.
A softer way is 60° for two hours in the oven.

Gudrun

-Ursprüngliche Nachricht-
Von: Campbell, Tasha M. via Histonet
[mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Mittwoch, 30. Mai 2018 14:15
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Masson's Trichrome Troubleshooting

Hello all,

I am having issues with my trichrome stain and I am about to lose my mind!
We just started doing it in house (although at my previous job I had done
trichrome by hand for years so I am not a stranger to it. And I never had
issues with it).  When I brought it in house at my current lab, I ordered
the same kit that I was familiar with.  Its PolySci.  I did the stain about
5 or 6 times and then all the sudden it quit working.  There was red where
it should be blue.  And there was blue staining but it was all in the
crypts.   I tried tweaking the stain a few times and nothing worked so I got
a new kit.  The first 2 times I used the new kit, it worked perfect!  But
after that it is back to doing the same thing again!  The collagen is not
staining blue.  It is staining red.  Can anyone please tell me why this is
happening?  I never had this issue before!  Thanks in advance!! See my
protocol below.

1. Mordant in Bouin's solution, microwave 5 minutes, allow to stand 15
minutes.
 2. Wash in running tap water to remove the picric acid, 5 minutes.
 3. Weigert's Working Hematoxylin, 10 minutes.
 4. Blue in running tap water for 5 minutes, rinse in distilled water.
 5. Biebrich scarlet for 5 minutes.
 6. Rinse in distilled water.
 7. Phosphotungstic/phosphomolybdic acid for 10 minutes.
 8. Transfer directly into Aniline blue for 5 minutes.
 9. Rinse in distilled water.
10. 1% Acetic acid for 1 minute.
11. Quick rinse and air dry.
12. Coverslip




Tasha Campbell, B.S.,HTL(ASCP)
Frederick Gastroenterology Associates
310 W. 9th St.
Frederick, MD 21701
301-695-6800 ext. 144

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] new developments in histopathology?

[Gudrun Lang via Histonet]

Dear histonetters,

I have to do financial planning for our diagnostic histolab until 2023 for
the necessitiy of new instruments etc.

As far as diagnostic development is a rather fast thing, I hardly can
imagine what will aproach us in 5 years in the clinical set.

 

What do you think? Which emerging technologies, besides the whole
molecularpathology, will be used in histomorphology. (in situ PCR?
Mutation-specific-in situ PCR?, automated FISH, .)

 

Many thanks for your inputs!

 

Gudrun Lang

 

(Linz, Austria)

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Modified GMS Protocols

[Gudrun Lang via Histonet]

Biological stains were mainly found by trial and error through the century. I 
think the main goals were to find a good result (for the purpose) within the 
available resources (cheap / expensive, easy/difficult to get) and with a 
practical application. 
Later also health-concerns played a role, that eliminated some protocols. I 
believe, that the "forefathers" had a hard time to find the best-working 
protocols. And after that, the histological community acts upon the sentence 
"never change a winning team" and sticks to the protocols. And we have to 
admit, that the chemical knowledge of the histologists and the histological 
knowledge of the chemists may be rather decreased than grown bigger. (a lack of 
universal scientists)

For the diverse oxidants I think (without literature as evidence) it is also a 
practical matter. The oxidizising result depends on strength of acid and 
duration of incubation. If you use a very strong oxidant, the time has to be 
watched very carefully and there is the risk of overoxidation. If you use a 
weaker oxidant, you have a longer time-space for a sufficient result. (5 min in 
periodic acid may refer to 20 sek in potassiumpermanganat ?) 

Gudrun



-Ursprüngliche Nachricht-
Von: Nipuna Weerasinghe via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Donnerstag, 28. Dezember 2017 20:30
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Modified GMS Protocols

I would like to know from subject matter expert why KMno4 never used as an 
alternative oxidant to CrO3 in Modified GMS. Oxidation of 1,2-glycol linkages 
in carbohydrates to aldehyde groups can be done by KMNO4 and used to do the 
same in Castella’s potassium permanganate-Schiff reaction, and Gordon and 
Sweets' reticulum. Moreover, KMNO4 is a strong oxidant that can oxidized 
aldehyde to carboxylic, so this leads to closer mimicking of function of CrO3.

Also why people only tried periodic and not any other oxidant to replace CrO3. 
I could not find any primary literates concnering this matter. Only handful of 
attempts with periodic is there.

Thanks for your answers in advanced

Lip.
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] gross photography

[Gudrun Lang via Histonet]

Hi,
We use MakroPath from Milestone in a routine histolab. The camera is mounted
on the top oft he grossing-station, with an integrated PC+monitor and pedals
for zooming and taking photos. Within this system you can mark the pictures,
draw something, measure something ...
In comparison to the older method with digital-camera, manual zoom etc. it
is very conveniant.  Picture quality is high.

Gudrun

-Ursprüngliche Nachricht-
Von: Julio Benavides Silván via Histonet
[mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Dienstag, 28. November 2017 21:27
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] gross photography


Hi there,

May I ask you your opinion about which system you are using to take gross
pictures? We are using a couple of big tungsten light bulbs and a Nikon d60
camera. We are a research lab working with sheep, so we get big lesions in
big organs. I was wondering if anybody is using a Digital Gross Photography
System and how they compare with a "more ytraditional" digital camera
approach.

As always, thank you so much for your opinions. Greatly appreciated!

Cheers

Julio





___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Ventana special stainer

[Gudrun Lang via Histonet]

Hello!

I have a question for those, who have experience with the "old" Ventana
special stainer Nexes and switched to the "new" special stainer Benchmark.
Is there a difference in the quality of the stainings? Better reagenses?
More possibilities to adapt the protocols?

Or is it just the same and only the deparaffination is now on board?

 

Thanks in advance

Gudrun Lang

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] decal protocol with EDTA for bonemarrow biopsies

[Gudrun Lang via Histonet]

Dear histonetters!

I would be happy about some input about decalcification protocols with EDTA
of trephine bonemarrow biopsies.

recommended duration of fixation?

recommended duration of decalcification?

strategies for speed-up of decal?

recommended EDTA-solution formula?

 

Hopefully some experienced histotechs can share their knowledge with me.

thanks in advance

Gudrun

 

 

 

 

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Digital Pathology & Coverslipping

[Gudrun Lang via Histonet]

Hi!
We scan our daily HEs for 1,5 year now. We have a Leica glass-coverslipper.
The brilliance of the images is very good. With film we had the experience,
that the brilliance of digital photos were never of the same quality as with
glass. (and had always issues with coming off the film after a year of
storage. So I doubt, that scanning older slides is successful.)
The drawback of the glass coverslipper is, that you have always to adjust
it. One day it is perfect without any bubbles, the other day the medium has
changed the consistency and you need more or less of it. When the medium
squeezes out a little bit, the scanner may recognise it as tissue. The same
happens, when the medium is spared at the edges and air goes in.
As solvent we use butyl-acetate, that evaporates very quickly and clean. The
slides are dry within minutes.
Our workflow is first to organise the cases for delivering, then scan the
sorted cases and then deliver the slides to the pathologists. So the slides
have enough time to dry and we can check the quality of coverslipping and
quality of the slide-barcode.

bye
Gudrun

-Ursprüngliche Nachricht-
Von: Haley Huggins via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Montag, 14. August 2017 17:48
An: Alexis Templeton
Cc: histonet@lists.utsouthwestern.edu
Betreff: Re: [Histonet] Digital Pathology & Coverslipping

Glass or film coverslips are fine, but you have to make sure they are clean,
no excess mounting media, or bubbles. The scanners pick up a lot of extra
things you don't want scanned. Also, check with your pathologists to see if
they have an opinion one way or another about which coverslips they prefer.

*Haley Huggins, HT (ASCP)cm*
*Technical Lab Supervisor*
*1050 Las Tablas Rd, Suite 14*
*Templeton, CA 93465*
*Office: 877-230-1518*
*Cell: 303-652-7453*

On Fri, Aug 11, 2017 at 12:42 PM, Alexis Templeton via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Hi All!
>
> My lab is considering moving up in the world of technology.  The goal 
> is to start scanning slides for pathologists to read digitally.  We 
> are a relatively high throughput lab and I'm trying to figure out what 
> we need in terms of an automatic coverslipper to avoid drying time.  
> We currently still coverslip by hand and I'm assuming there would be 
> too much wet glue to place the slides directly in a scanner.  Tips and 
> recommendations, please!
>
> Alexis Templeton, HT (ASCP)CM
> Diagnostic Laboratory Supervisor
> Histopathology
> Texas A Veterinary Medical Diagnostic Laboratory P.O. Drawer 3040 | 
> College Station, TX 77841-3040
> p: (979) 845-3414 | f: (979) 845-1794
> atemple...@tvmdl.tamu.edu
> http://tvmdl.tamu.edu
>
> We Moved!  Effective February 27, 2017 our physical (shipping) address 
> is
> 483 Agronomy Road, College Station, TX  77840. Our billing address 
> remains at PO Drawer 3040, College Station, Texas  77841  
> **The contents of this email do not necessarily represent the views or 
> policies of TVMDL. This email is intended for the recipient only and 
> the information should not be released to third parties.
>
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Tissue Fixation

[Gudrun Lang via Histonet]

I second the opinion of Joyce. We see such effects in portio-conisations, that 
are done with a thermo-electrical knife. The surface and the underlying area 
show a very pink colour in HE. It can also be seen in prostata-chips. IHC on 
such biopsies shows the effect of an non-stainable edge with a clear cut 
between positive and negative. 
https://www.uni-marburg.de/fb20/zahnerhaltk/lehre/Download/Bilder/35_plattenepithelmetaplasie_portio_02212427.jpg
I've found this picture, that shows a typical conisation-section.
regards
Gudrun 

-Ursprüngliche Nachricht-
Von: Weems, Joyce K. via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Freitag, 31. März 2017 17:26
An: Morken, Timothy; Rene J Buesa; T H; Histonet@lists.utsouthwestern.edu
Betreff: Re: [Histonet] Tissue Fixation

Aren't LEEPS done with some sort of electric method that will damage the tissue 
before it even reaches formalin. I'm not positive but Google it - I believe 
that might be the problem. j

Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
770-380-8099 Cell
joyce.we...@emoryhealthcare.org



www.saintjosephsatlanta.org
5665 Peachtree Dunwoody Road
Atlanta, GA 30342

This e-mail, including any attachments is the property of Saint Joseph’s 
Hospital and is intended for the sole use of the intended recipient(s).  It may 
contain information that is privileged and confidential.  Any unauthorized 
review, use, disclosure, or distribution is prohibited. If you are not the 
intended recipient, please delete this message, and reply to the sender 
regarding the error in a separate email.

-Original Message-
From: Morken, Timothy via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Friday, March 31, 2017 11:07 AM
To: Rene J Buesa ; T H 
Cc: Histonet 
Subject: Re: [Histonet] Tissue Fixation

Tim, I have to agree with Rene that the formalin or time in formalin is 
obviously not the problem - it has plenty of time in formalin (and who would 
dilute it anyway?). Handling before formalin must always be determined when 
problems arise. If the sample sits on a paper towel, gauze etc it does not 
really degrade faster, rather the tissue may dry out and so fixes by 
dehydration rather than by formalin. It may be the formalin cannot get into the 
tissue in those dried out portions of the tissue. However that is just 
speculation. Since these all seem to be from one particular client, the client 
is the place to start. The only way to determine the problem is to follow the 
specimen from start to finish. Can you or someone you trust physically observe 
the way samples are handled from the time they are taken to the time put in 
formalin? One issue I always run up against is people  saying they do one thing 
but actually doing another. And they may realize during questioning that they 
are not doing it right but don't want to admit it. It wastes a lot of time. 
I've had physicians tell me to ignore the part of the process they are 
responsible for because they do it right. I just tell them that to be complete 
we need to follow the process from start to finish, nothing personal, just 
business. Leaving any part out may lead to not resolving the problem. Probably 
99% of the process is just fine, but that 1% is damaging the sample and needs 
to be illuminated.


Tim Morken
Pathology Site Manager, Parnassus
Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of 
Pathology UC San Francisco Medical Center



-Original Message-
From: Rene J Buesa via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Friday, March 31, 2017 6:33 AM
To: T H; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Tissue Fixation

What you describe as a possible scenario is absolutely possible.If your PT does 
not "want to hear" about it, suggest she gets a "hearing aid" or to study 
something about histotechnology or even better yet, pay attention to what a 
professional on the subject (you) has to say about it. You would never dare to 
question her diagnosis, why would she question yours on this subject?René

On Friday, March 31, 2017 9:11 AM, T H via Histonet 
 wrote:


 Good Morning,


I have a pathologist that is not happy with the fixation on some of our LEEP 
specimens.  She swears its histology doing something to the specimen to cause 
the tissue to look unfixed on only "part" of the LEEP specimens (all the same 
client specimens).  She claims we must be diluting our formalin to cause this 
issue or "something".  We mentioned maybe it was on the clients end not placing 
them in 10% formalin right away, she wouldn't hear of it.


Let me give you some back ground on how our process works.  Our clients send us 
all our specimens to us via Overnight FedEx or UPS in 10% formalin they will 
then they sit in 10% formalin in-house until the processors are started around 
3pm and sits 

[Histonet] WG: question about retina IHC

[Gudrun Lang via Histonet]

Because I have no experience with this special specimens, I give the
question below to the kind histonet and its professionals.

Gudrun Lang

 

Von: Amirkavei Mooud [mailto:mooud.amirka...@aalto.fi] 
Gesendet: Donnerstag, 23. März 2017 13:02
An: Lang Gudrun
Betreff: question about retina IHC
Wichtigkeit: Hoch

 

Dear Gurdun,

 

I am a Ph.D student from Kai Kaarniranta’s grou, Finland. I am working with
mouse retina and need to do retina staining to check desired changes in POS.
Now I would like to ask your help. Would it be possible for you to help me
with the protocol of preparation of sample for retina sectioning and all
procedure of its IHC. I don’t know if I can use snap freezing and
cryosectioning or using PFA and normal microtome?

 

I appreciate very much your kind help J.

 

Bets regards,

Mooud

 

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] staining samples years in formalin

[Gudrun Lang via Histonet]

Hi Mary Lou,
I think the problem is, that proteoglycanes will be solved by acids through
decal. Try EDTA decal or prolong the staining time in Alcianblue - maybe for
2 hours?
Try to decal your controls in the same manner and compare the results.
I don't think, that formalin fixation is the culprit, but long storage in
aequous solutions like formalin may influence watersoluble proteoglycans.

Gudrun

-Ursprüngliche Nachricht-
Von: Mary Lou Norman via Histonet [mailto:histonet@lists.utsouthwestern.edu]

Gesendet: Mittwoch, 18. Januar 2017 15:01
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] staining samples years in formalin

Dear Histonetters,
My samples have been in formalin up to 3 years and there have been
complaints about poor staining. Please help me with solutions if there are
any. My controls have not been in formalin as long. The complaint has been
with the Alcian Blue pH2.5 and Saf O specifically.
The samples are stifles so I need to decal them also. I use formic/na
citrate.
Any and all comments, suggestions are much appreciated. Thank you.
Mary Lou Norman

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Questions RE: "what I have found about p16" and cellblocks for controls

[Gudrun Lang via Histonet]

Your concerns are reasonable. Cyto-specimens are usually fixed in alcoholic
solutions not in NBF. Alcohol-fixation gives false positives in Her2. This
can also be seen in underfixed tissue, that is mainly fixed by the ethanols
in the processor.

Her2-protein is a normal protein on the cellsurface. FFPE treatment "turns
it down" and normal amount cannot be detected. This is "negative". Only Her2
positives have abundant protein to be detected with our methods. That is why
correct standardizised treatment is important to avoid "false positives" and
"false negatives".

Gudrun
leading histotechnologist,
Kepler Universitätsklinikum Linz
Austria

-Ursprüngliche Nachricht-
Von: Cassie P. Davis via Histonet [mailto:histonet@lists.utsouthwestern.edu]

Gesendet: Freitag, 6. Januar 2017 18:26
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Questions RE: "what I have found about p16" and
cellblocks for controls



Is there anyone in Histonet land who is using both the Optiview and
Ultraview on the XT and ULTRA at the same time? I'd like to ask a few
questions, please email me.



We have a cytology cellblock case that came out stunningly positive for Her2
and one of our pathologist thought it would make a dynamic control if there
were any specimen left to make the control block.

While I am excited about the possiblity (Her2 control is hard to come by
around here) some concern is expressed about using it because it is initally
fixed in cytolyt before FFPE. I can argue both sides of this. I would enjoy
your input on this folks.


Cassandra Davis
Histology Technician
AP Laboratory
302-575-8095
Email:  cda...@che-east.org



From: Cassie P. Davis
Sent: Thursday, January 05, 2017 10:02 AM
To: histonet@lists.utsouthwestern.edu
Subject: what I have found about p16


Hi Histonet folks,

 Thank you for all the help, for those who are following what I
have found:

(1)



Sigma-Aldrich does have p16 (it is also know as INKa):



Anti-INKa(p16) antibody, clone 13H4.1   cat # MABE1328

or

Anti-p16 Antibody, clone D25  cat#MAB4133



(2)



Stacy was kind enough to share this:





Hi Cassie,
Are you referring to Ventana?
If so, they will have 2 other kits coming out for it.  They will require the
use of Optiview detection or it will be considered an LDT.  Just found this
out from my rep.
Thanks,
Stacy

Stacy McLaughlin, HT (ASCP)
Histology Supervisor
Cooley Dickinson Healthcare
30 Locust Street
Northampton, MA 01060
(413-582-2019



Stacy, thanks for sharing! We did not get that information from our rep.


Cassandra Davis
Histology Technician
AP Laboratory
302-575-8095
Email:  cda...@che-east.org


Confidentiality Notice:
This e-mail, including any attachments is the property of Trinity Health and
is intended for the sole use of the intended recipient(s). It may contain
information that is privileged and confidential.  Any unauthorized review,
use, disclosure, or distribution is prohibited. If you are not the intended
recipient, please delete this message, and reply to the sender regarding the
error in a separate email.
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Staining stomach cells

[Gudrun Lang via Histonet]

Did you look at the mucin-antibodies? 
Pepsinogen-IHC should be positiv in chief-cells.
Gastrin-IHC should be positiv in parietal-cells.
Chromogranin A should be positiv in endocrine cells.

just my ideas. please confirm with your own research. 

Gudrun

-Ursprüngliche Nachricht-
Von: Judi Ford via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Mittwoch, 15. Juni 2016 22:16
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Staining stomach cells

Hi everyone,
I'm trying to differentiate parietal vs chief vs endocrine cells in the
stomach. Any ideas on stains or antibodies I could use.
Thanks,
judi
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] PAS/Decal Question

[Gudrun Lang via Histonet]

Hi Pam,
my personal opinion is, that 2 hours fixation is too short for sufficient
tissue-protection before acid decalcification. Formic acid at 50°C must have
an impact on glycoproteins. Wether it is a kind of solving the "sugars" or
beginning oxidation of the OH-groups (like periodic acid does in the
PAS-reaction).

In our lab we do also acid decal with formic acid for at least 6 hours at
RT, after one day in NBF. Our processing protocol is the routine-protocol
over night. How thick are your BMT, also 3-4 mm?
In my opinion 4 hours are a challenge. Are the other stainings of the BMT
optimal or show sometimes similar outcome? "Smudginess" reminds me of
insufficient infiltration.

I also see that our PAS is not as bright as in the other specimens without
decal. Sometimes it gives more the impression of a diastase-PAS. 

Gudrun

-Ursprüngliche Nachricht-
Von: Marcum, Pamela A via Histonet
[mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Dienstag, 03. Mai 2016 18:39
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] PAS/Decal Question

We are still having issues with our PAS stain on decaled bone marrows.  The
Pathologists in HemePath are seeing what they refer to as smudginess in
cells on some areas of the completed PAS slides.  We have looked at
everything and cannot find where the issue is coming from at this point.  We
have done manual staining for PAS, automated on the Leica stainer and on the
Dako Artisan.  All methods show the same result for some slides.  We can go
for several days to a week or more with no problem and then suddenly it is
back and we have changed nothing in the way we do the processing, embedding,
sectioning, deparaffinization and coverslipping.  We do as many as 38 bone
marrow cores a night or as few as 8 and can find no correlation in the
number we have to deal with for a given period.  All bone marrows drawn
today must be completed by 8AM tomorrow morning.

Fixation after pulling the bone marrows is a minimum of 2 hours in AZF with
a maximum of 7 hours +/-.

Grossed and placed in cassettes for 15 minute rinse in running DI Water

Decal currently in the Milestone Decal Unit for 45 minutes in Immunocal at
50C

Rinsed in running DI water for 15 minutes

Placed in 10% NBF and processed on a 4 hour program with a delay of 4 hours
minimum to come off at 4:45AM.

If anyone knows of any literature on decal effects on PAS staining in bone
marrows please contact me.  This has been going on for months and no matter
what we do manual staining, Leica adaptation for automated or Dako it is not
helping.  Dako has been great with sending in technical experts repeatedly
and we cannot get this corrected.

Thanks,
Pam

--
Confidentiality Notice: This e-mail message, including any attachments, is
for the sole use of the intended recipient(s) and may contain confidential
and privileged information. Any unauthorized review, use, disclosure or
distribution is prohibited. If you are not the intended recipient, please
contact the sender by reply e-mail and destroy all copies of the original
message.
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] PAS Stain

[Gudrun Lang via Histonet]

As far as I remember the incubation temperature is at roomtemperature. 60°C
would rather denature the native enzyme than increase activity.

Look at the optimal working temp of the reagens you have bought.

regards
Gudrun

-Ursprüngliche Nachricht-
Von: Joanne Clark via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Mittwoch, 04. Mai 2016 22:03
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] PAS Stain

Hi Histonetters, I'm hoping someone can help me troubleshoot our PAS
diastase method.  We have been digesting the tissue in 0.5% diastase of malt
in a 60 degree oven for 30 minutes, but do not see the glycogen being
digested out.  I have tried alpha amylase and beta amylase also without any
luck.  Does anyone have any suggestions to get the digestion to work

Joanne Clark, BAAS, HT(ASCP)CM
Director of Histology

P.   (575) 622-5600
C.   (575) 317-6403
F.   (575) 622-3720
TF. (800) 753-7284

pcnm.com




Disclaimer: This electronic message may contain information that is
proprietary, confidential, or legally privileged or protected. It is
intended only for the use of the individual(s) and entity named in the
message. If you are not an intended recipient of this message, please notify
the sender immediately and delete the material from your computer. Do not
deliver, distribute or copy this message and do not disclose its contents or
take any action in reliance on the information it contains.
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] WG: copper stain

[Gudrun Lang via Histonet]

Thank you all for your responses.
Gudrun

-Ursprüngliche Nachricht-
Von: Gudrun Lang via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Mittwoch, 20. April 2016 18:42
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] copper stain

Hi all!

Which stain would you prefer to demonstrate copper? Rhodanin or Victoria
blue?

 

Thanks in advance

Gudrun

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] copper stain

[Gudrun Lang via Histonet]

Hi all!

Which stain would you prefer to demonstrate copper? Rhodanin or Victoria blue?

 

Thanks in advance

Gudrun

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] formalin and shrinkage

[Gudrun Lang via Histonet]

My question refers to the difference in dimensions of the native tissue and the 
fixed tissue. – so without any  influence of ethanol and xylol.

 

I have no access to the whole publication you mentioned. If I understand the 
summary correctly, 2,5% shrinkage was found after formalinfixation.

 

There is no actual problem to solve, just academic question. 

Gudrun

 

 

Von: Jay Lundgren [mailto:jaylundg...@gmail.com] 
Gesendet: Montag, 22. Februar 2016 20:31
An: gu.l...@gmx.at
Cc: histonet
Betreff: Re: [Histonet] formalin and shrinkage

 

I was taught at AFIP to expect shrinkage of 10%, in each dimension.  So I guess 
that's 30% shrinkage overall?  Shrinkage is partially caused by formalin 
crosslinking the proteins in fixation, and partially by dehydration.  Maybe a 
little shrinkage in xylene too?  From removal of fat in adipose tissues?  

http://link.springer.com/article/10.1007/BF00695061#page-1

 

Is your Pathologist really concerned about shrinkage, or about curling and 
distortion of small shave bxs?  Because a certain degree of shrinkage is an 
unavoidable artifact of tissue processing.

 

If it's the latter, I like to use 2 blue sponges.  I find they really help to 
keep things flat and oriented.  Some people don't like them because of 
carryover.  I just say change your processor reagents more often.

 

Sincerely,

 

Jay A. Lundgren, M.S., HTL (ASCP) 

 

 

On Mon, Feb 22, 2016 at 9:59 AM, Gudrun Lang via Histonet 
<histonet@lists.utsouthwestern.edu> wrote:

Hi!

Today someone asked me about shrinkage caused by the fixation with
formaldehyde specially on skin-biopsies.  She spoke about shrinkage of 30%
percent. In my opinion shrinkage is mainly caused by the processing with
dehydration and defatting. Formaldehyde renders the tissue harder but not
strictly smaller.



What is the opinion of the community?



Gudrun







___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

 

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Davidson's fixative and IHC question

[Gudrun Lang via Histonet]

I think fixing with Davidson is ethanol-fixation in the first line. There
are some antigens, that are susceptible to ethanol. 

Gudrun

-Ursprüngliche Nachricht-
Von: Judi Ford via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Montag, 22. Februar 2016 18:45
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Davidson's fixative and IHC question

Hi Everyone. Hope you all had a really good weekend. Thanks for the replies
to my double ihc question.

I do have another question; this one is from a friend of mine. Her client
wants to do ihc on rabbit eye tissue. The tissue will be fixed in Davidson's
fixative for 24 hours then in 10% NBF. Will this have affect future ihc
projects?

Thanks,
Judi
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] formalin and shrinkage

[Gudrun Lang via Histonet]

Hi!

Today someone asked me about shrinkage caused by the fixation with
formaldehyde specially on skin-biopsies.  She spoke about shrinkage of 30%
percent. In my opinion shrinkage is mainly caused by the processing with
dehydration and defatting. Formaldehyde renders the tissue harder but not
strictly smaller. 

 

What is the opinion of the community?

 

Gudrun

 

 

 

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Double stain IHC question

[Gudrun Lang via Histonet]

In my opinion, this would only be possible, if the commercial and the
homegrown antibody are from different species. For example one from mouse
and one from rabbit.
Then you can proceed with different secondaries (goat anti mouse conjugated
with peroxidase, goat anti rabbit conjugated with alkaline phosphatase).
Then chromogens that work with each of the enzymes. 

If the antibodies are from the same species I see no way to distinguish
both. Only if one is conjugated with biotin and the other with digoxigenin,
then you could proceed with secondaries against biotin and digoxigenin.
etc..

Gudrun

-Ursprüngliche Nachricht-
Von: Judi Ford via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Samstag, 20. Februar 2016 01:54
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Double stain IHC question

Hi everyone,
I have a question in chromogenic double staining. Here is the situation.
Tissue = human, frozen
Antibody = same protein (A)

1.   Commercial antibody of A

2.   Homegrown antibody of A, human, biotinylated

Question: can you stain both versions of this antibody on the same tissue,
same slide? Goal is to see where each stains in the tissue and if they
co-localize. If they do co-localize then how do you distinguish between that
and where they stain individually? Would you use different chromogens and
hope that where they come together it turns a different color?
I am really interested if this can work. Thanks in advance for any replies.
Judi
South San Francisco, CA
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] keratohyalin granules

[Gudrun Lang via Histonet]

Do you stain with hematoxylin only? or also with eosin. I think eosin is the
dye, that would highlight the basic proteins in the granula.
Gudrun

-Ursprüngliche Nachricht-
Von: Kalleberg, Kristopher via Histonet
[mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Mittwoch, 17. Februar 2016 21:37
An: histonet@lists.utsouthwestern.edu;
histonet-requ...@lists.utsouthwestern.edu
Betreff: [Histonet] keratohyalin granules

Hello All,

Does anyone have experience staining the keratohyalin granules in skin with
different types of hematoxylins?  I am currently looking at keratohyalin
granules and staining with Harris hematoxylin and there seems to be little
staining and not as prominent as I would have expected.  Wonder if anyone
has noticed that different hematoxlins stain keratohyalin granules better
than others.  The skin is normal skin photoprotected and photoexposed.
Thank you in advance for any helpful insights.

Kris
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Sponges for processing biopsies

[Gudrun Lang via Histonet]

Sponges are available in different qualities. We use very soft ones, that
don't hurt the tissue. 
I know, there are also very hard materials on the market, that may render
holes in the underfixed biopsies.

Gudrun

-Ursprüngliche Nachricht-
Von: Lester Raff MD via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Freitag, 29. Jänner 2016 20:05
An: 'histonet@lists.utsouthwestern.edu'
Betreff: [Histonet] Sponges for processing biopsies

WE use the biopsy sponges, THOROUGHLY soaked in formalin. They are easy to
use and not time consuming either for the grosser or the histologist. About
98% of our volume is prostate biopsies, and we do not see the compression
artifact Rene references.





Unrelated blog http://tinyurl.com/down0129


A good weekend to all.

Lester J. Raff, MD MBA
UroPartners
Medical Director Of Laboratory
2225 Enterprise Dr. Suite 2511
Westchester, Il 60154
Tel: 708-486-0076
Fax: 708-492-0203

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Nuclear bubbling artifact

[Gudrun Lang via Histonet]

I've read about a group, that observed living cells during the
fixation-process. They saw bubbling in the first period of contact and
penetration of formaldehyde. After a certain time the bubbles disappeared
again. 
Along this observation for me bubbles are a sign of too short fixation.

Gudrun Lang

-Ursprüngliche Nachricht-
Von: Teri Johnson via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Freitag, 29. Jänner 2016 19:49
An: 'histonet@lists.utsouthwestern.edu'
Betreff: Re: [Histonet] Nuclear bubbling artifact

Hi James,

Nuclear bubbling artifact is most commonly seen in formalin fixed epithelial
cells, and GI biopsies are among those samples that are particularly
susceptible to it. It has been linked to inadequate fixation and also to
heating of slides prior to staining without complete air-drying of the
tissues. I would recommend cutting the block again, air drying the slides
for a time before using heat to melt the wax prior to H stain and see if
the artifact persists.

http://www.cap.org/apps/docs/proficiency_testing/nuclear_bubbling.pdf

Best wishes,
Teri Johnson




CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is
for the sole use of the intended recipient(s) and contains information that
is confidential and proprietary to Genoptix Medical Laboratory or its
subsidiaries. Any unauthorized review, use, disclosure or distribution is
prohibited. If you are not the intended recipient, immediately contact the
sender by e-mail and destroy all copies of the original message.
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] additional barcode on ventana-labels

[Gudrun Lang via Histonet]

Hi!

Did anyone manage to bring Ventana to open their label-design? We need an
additional 2D-Barcode with the slide-data on the label.

 

Regards

Gudrun Lang

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Hirschsprung diagnostic kit

[Gudrun Lang via Histonet]

Hi all!

Has anyone experiences with the Hirschsprung diagnostic kit from Bio-Optica
(Italian company)? This kit contains lyophilizised reagenses, that have to
be restored before use. It comprises the detection of AChE, SDH, ANE and
NADPH.

 

Any input is appreciated.

regards

 

Gudrun Lang

histotech, Linz Austria

 

 

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] processing cycle

[Gudrun Lang via Histonet]

Hi Allison,
we doubled the times of the regular processing protocol beginning with
longer absolute ethanol, intermedium and paraffin. Our regular protocol
takes 13 hours and the fatty-protocol takes 17 hours. We start it at about 2
pm with endtime at 8 am. So breast tissue is mostly embedded at the end of
the bulk of cassettes.

This improved cutting really in a great manner.  The problem occurs, that
fixation time is rather short, when grossing of breast is done just shortly
before processing. So we want our pathologists to gross the breast from the
day before rather early in the morning. 

Gudrun

-Ursprüngliche Nachricht-
Von: Eck, Allison via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Dienstag, 17. November 2015 20:15
An: 'histonet@lists.utsouthwestern.edu'
Betreff: [Histonet] processing cycle

Does anyone have a processing cycle that is good for fatty tissues like
breast that they would be willing to share?  This will be used on a VIP5.

Thank you in advance

Allison
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Eosinophil labelling

[Gudrun Lang via Histonet]

What about simple H?
Gudrun

-Ursprüngliche Nachricht-
Von: Julio Benavides via Histonet [mailto:histonet@lists.utsouthwestern.edu]

Gesendet: Montag, 14. September 2015 18:52
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Eosinophil labelling

Hi everyone,

Is there anti antibody , working on FFPE, labelling eosinophils? if it works
in sheep then that´s perfect!

I have seen several histochemical methods (Modified Congo Red, Luna
Protocol, Astra Blue/Vital New Red Protocol (AB/VNR, Sirius Red Stai ). 
Does anyone has any experience with them?

Any help/comments in this issues would be, as always, greatly appreciated!!!

Thank you

Thanks a lot!

Julio

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Pre-floating tissue sections in dilute alcohol

[Gudrun Lang]

Some of our histotechs use just cold tapwater for wrinkled sections before
transferring them  into the warm waterbath. I don't know, if the added
ethanol makes any difference (maybe reducing surface tension).

I would offer the possibility of using the additional waterbath, but let the
histotechs decide, if it is necessary to use it.

Gudrun Lang
Leading histotech, Linz Austria


-Ursprüngliche Nachricht-
Von: Garrey Faller [mailto:garr...@gmail.com] 
Gesendet: Samstag, 06. Juni 2015 16:29
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Pre-floating tissue sections in dilute alcohol

Hi everyone,

I just became aware of this technique last week, and it seems to work great.
I did a quick google search and found this quick reference.
http://www.scielo.cl/pdf/ijmorphol/v30n1/art07.pdf
http://www.scielo.cl/pdf/ijmorphol/v30n1/art07.pdf
Anyone out there float their sections in dilute ethyl alcohol before
transferring to the water bath?
Its an extra step and introduces an extra chance to introduce floaters.
But, the quality seems to be improved.
Any thoughts?  

Thanks in advance.
Garrey Faller
Pathologist
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] FW: Masson's trichrome stain

[Gudrun Lang]

Maybe I am wrong, but I think the usage of plastic forceps for trichrome
stains is nonsense. I only know this from silver stains, but also have never
seen problems personally.

The polyacids have to stay onto the section, when anilinblue is added. There
exists a theory, that PMA/PTA acts like a mordant and builds bridges between
collagen and anilinblue. It is proven, that PMA is not washed out of the
tissue and is bound to the fibers. On the other hand it seems, that the
polyacids prevent the staining of globular proteins by anilinblue (the
longer the polyacids the brighter the red colour).
It also helps to keep the low staining-pH of anilinblue.

Gudrun Lang




From: Justine Lanzon [mailto:justinelan...@hotmail.com]
Sent: Thursday, March 05, 2015 5:36 AM
To: lindamarg...@gmail.com
Subject: Masson's trichrome stain

 

Hi,

I am doing a write up on Masson's trichrome stain however I cannot answer
these two questions:

- Why are plastic forceps used instead of metal ones to hold the stained
slide?

- Why do we not rinse before Alinine blue?

 

Can you please help me?

 

Many Thanks,

Justine Lanzon

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] Stain vs dye and control

[Gudrun Lang]

Hi Jorge,
Not all histolabs have access to fresh cultures. So they search an easy way to 
get bacterial controls. And things like sausage are more like the usual 
specimens than liquid samples.

The best staining control is an inhouse-specimen, that is processed in the same 
way as any other specimen (preanalytic, fixation, processing, cutting, 
staining). But sometimes this is not possible, so one uses the next-best.
A control with known ingredients (like bacteria) can be used to check the whole 
process and must be positive for the tested parameter. A patient-sample can 
only be considered positive or negative, if the positive-control proves the 
functionality of the process (staining-protocol).

Dye and stain. You can touch the dye, but not the stain.  And then you have got 
stained fingers. ;-)

Gudrun



-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Jorge A. 
Santiago-Blay
Gesendet: Freitag, 06. März 2015 21:49
An: Histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Stain vs dye and control

Dear Histonetters:

Last semester I taught a microbiology lab and, as I was reviewing for class, 
noticed some lack of precision in the use of the terms dye vs.
stain in biology. Could someone help?

While I am on stains, I have been following the emails on controls and wonder 
a couple of things.

1. What would testing for bacteria on beef jerky, hot dogs or burgers 
accomplish that is different (ideally better) that what one accomplishes by 
pulling out from fresh cultures out of a medium (e.g. liquid, such as broth, 
solid, such as slabs, agar)?  Is it the idea to test for bacteria in an animal 
tissue? If so, would a solid medium (like someone mentioned recently, such as 
agar) do?

2. An advantage of using fresh bacterial cultures of known Gram is that it 
could be used to test whether the reagents are good enough. Last semester I had 
the suspicion that one (or more) of our Gram reagents where not up to par.

If you have any feedback, please feel free to email me directly at 
blayjo...@gmail.com . Thank you.

Sincerely,

Jorge

Jorge A. Santiago-Blay, PhD
blaypublishers.com
http://blayjorge.wordpress.com/
http://paleobiology.si.edu/staff/individuals/santiagoblay.cfm
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Considerations about Warthin-Starry principle

[Gudrun Lang]

Dear histonetters,

 

I came across a question regarding the Warthin-Starry principle. Are
bacterial catalase or oxidase the cause for developement of silver-deposits?

 

The usual explanation is, that bacteria are able to bind silver onto their
surface, that is further reduced by hydrochinon to visible silver-deposits.
But why?

Some considerations:

The pH must be held at pH4 with citric acid in the WS developement-solution.
(Is this the pH-optimum for the bacterial enzymes?)

IHC shows that endogenous peroxidase survives the embedding-process. (Is
this also true for the bacterial enzymes?)

Spirochetes (and others) are catalase and oxidase positive.

Silver-In situ hybdridisation technique shows, that peroxidase with H2O2 can
be used to reduce Silveracetate for silver-deposits. 

 

Is there a possible relationship between the bacterial enzymes and the WS
principle?

Or am I totally on an erroneous way? What do you think?

 

thanks for any input.

Gudrun Lang

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] Trichrome Fixation

[Gudrun Lang]

I disagree. Usually a dye binds covalently to the mordant, and a mordant should 
be a multi-valent metal (like aluminium, ferrum, molybden,..). The 
dye-mordant-complex binds via the mordant to the substrate.
Picric acid of Bouin's is washed out before staining.

It may be a matter of definition of the term mordant.

Gudrun

-Ursprüngliche Nachricht-
Von: Johnson, Carole [mailto:cjohn...@nmda.nmsu.edu] 
Gesendet: Freitag, 06. März 2015 15:10
An: gu.l...@gmx.at
Betreff: RE: [Histonet] Trichrome  Fixation

Boiuns solution acts as a mordant in trichrome stains

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang
Sent: Friday, March 06, 2015 1:13 AM
To: 'Amos Brooks'
Cc: histonet@lists.utsouthwestern.edu
Subject: AW: [Histonet] Trichrome  Fixation

Hi,
years ago we did a stain called CAB (=one step trichrome) regularly on 
liver-tissue. I don't know if it was because of ignorance or with aim, but it 
was done without Bouin. The result was blue-grey hepatocytes and darker blue 
collagen.  - also totally different to the result with Bouin (red hepatocytes).

I think the Bouin is less a re-fixation than more an binding-site retrival 
in this context.

Gudrun


-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Amos Brooks
Gesendet: Donnerstag, 05. März 2015 22:01
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Trichrome  Fixation

Hi,
 It is interesting that you should mention the importance of fixation on 
the Trichrome stain. I have an image of two murine hearts processed, cut and 
stained side by side. The only difference between the two is that they were 
harvested at different times, so one sat in formalin long enough to be properly 
fixed the other one was placed in the fixative then immediately brought in to 
be processed from 70% ethanol on. They are *totally* different looking. The red 
muscle tissue looks more purple therefore less distinct from the blue blood 
vessels. You get a similar effect with lung bronchial epithelium.

Cheers,
Amos

On Tue, Mar 3, 2015 at 1:00 PM, histonet-requ...@lists.utsouthwestern.edu
wrote:

 Message: 15
 Date: Tue, 03 Mar 2015 12:34:33 -0500
 From: John Kiernan jkier...@uwo.ca
 Subject: Re: [Histonet] Masson trichrome and H and E
 To: Emily Brown talulahg...@gmail.com,
 histonet@lists.utsouthwestern.edu
 histonet@lists.utsouthwestern.edu
 Message-ID: 7390afa3112a.54f5a...@uwo.ca
 Content-Type: text/plain; CHARSET=US-ASCII

 If you can't get two colours with HE, don't expect to get the colour 
 scheme right with Masson's trichrome, which needs more skill. If you 
 are hoping to show basement membranes in the kidney, you would do 
 better to use a technically simpler staining method such as 
 picro-sirius red or periodic acid-Schiff. If for some reason you 
 really need three colours, a one-step trichrome such as Gomori's, 
 Cason's or Gabe's might be the way to go rather than Masson's or one 
 of the other multi-step trichromes. Remember that all trichrome 
 methods are greatly influenced by the fixative. A post-fixation 
 treatment of the sections, usually with picric acid, is needed for 
 formaldehyde-fixed tissues. Some alternative post-fixation treatments 
 were proposed by Yu  Chapman (2003) J. Histotechnol. 26(2): 131-134, but 
 their coloured photos were not very convincing.

 Making up staining solutions in-house is always cheaper than buying 
 pre-made solutions.

 John Kiernan
 London, Canada

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



Confidentiality Notice: New Mexico has a very broad public records law. Most 
written communications to or from state employees are public records. Your 
e-mail communications may therefore be subject to public disclosure. This 
e-mail, including all attachments is for the sole use of the intended 
recipients. Any unauthorized review, use, disclosure or distribution is 
prohibited unless specifically provided under the New Mexico Inspection of 
Public Records Act.


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] Trichrome Fixation

[Gudrun Lang]

Hi,
years ago we did a stain called CAB (=one step trichrome) regularly on 
liver-tissue. I don't know if it was because of ignorance or with aim, but it 
was done without Bouin. The result was blue-grey hepatocytes and darker blue 
collagen.  - also totally different to the result with Bouin (red hepatocytes).

I think the Bouin is less a re-fixation than more an binding-site retrival 
in this context.

Gudrun


-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Amos Brooks
Gesendet: Donnerstag, 05. März 2015 22:01
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Trichrome  Fixation

Hi,
 It is interesting that you should mention the importance of fixation on 
the Trichrome stain. I have an image of two murine hearts processed, cut and 
stained side by side. The only difference between the two is that they were 
harvested at different times, so one sat in formalin long enough to be properly 
fixed the other one was placed in the fixative then immediately brought in to 
be processed from 70% ethanol on. They are *totally* different looking. The red 
muscle tissue looks more purple therefore less distinct from the blue blood 
vessels. You get a similar effect with lung bronchial epithelium.

Cheers,
Amos

On Tue, Mar 3, 2015 at 1:00 PM, histonet-requ...@lists.utsouthwestern.edu
wrote:

 Message: 15
 Date: Tue, 03 Mar 2015 12:34:33 -0500
 From: John Kiernan jkier...@uwo.ca
 Subject: Re: [Histonet] Masson trichrome and H and E
 To: Emily Brown talulahg...@gmail.com,
 histonet@lists.utsouthwestern.edu
 histonet@lists.utsouthwestern.edu
 Message-ID: 7390afa3112a.54f5a...@uwo.ca
 Content-Type: text/plain; CHARSET=US-ASCII

 If you can't get two colours with HE, don't expect to get the colour 
 scheme right with Masson's trichrome, which needs more skill. If you 
 are hoping to show basement membranes in the kidney, you would do 
 better to use a technically simpler staining method such as 
 picro-sirius red or periodic acid-Schiff. If for some reason you 
 really need three colours, a one-step trichrome such as Gomori's, 
 Cason's or Gabe's might be the way to go rather than Masson's or one 
 of the other multi-step trichromes. Remember that all trichrome 
 methods are greatly influenced by the fixative. A post-fixation 
 treatment of the sections, usually with picric acid, is needed for 
 formaldehyde-fixed tissues. Some alternative post-fixation treatments 
 were proposed by Yu  Chapman (2003) J. Histotechnol. 26(2): 131-134, but 
 their coloured photos were not very convincing.

 Making up staining solutions in-house is always cheaper than buying 
 pre-made solutions.

 John Kiernan
 London, Canada

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] Masson trichrome and H and E

[Gudrun Lang]

Hi!
For lacking Eosin-staining check the pH of the solution. It should be 4-5.
With some drops of acetic acid you may recover the staining-action.
Gudrun
= = =
On 03/03/15, Emily Brown  talulahg...@gmail.com wrote:
 Hello!
 
 I am doing Masson trichrome manually (not with a machine) and I just 
 found out the kit is $800!! I was thinking of buying the ingredients 
 separately, but why the hell is it so expensive???
 Also, the people I was going to borrow their reagents from said their 
 aniline blue is not very good and I wanted to replace it. I only need 
 about 250ml, what brand do you guys prefer?
 l know I can google this, but I want to know what you guys like and 
 what works best. This is for mouse kidney paraffin sections, 4 to 5
microns.
 Another question, I did H and E and there is no eosin staining. I 
 think the reagents are pretty old, so I thought that might be a 
 problem. Also because my lab is cheap, they were reusing the xylenes 
 and EtOH for both rehydrating and rehydrating. I told my boss this is 
 probably not a good idea as the end steps will have stain in them. And 
 I also think this is why it didn't work! The EtOH is also really old 
 so who know is the 100% is actually even close to 100% any more. I'm 
 buying new reagents, but if you guys think anything else would help, let
me know.
 
 Also, shoutout to Ann, I know you're reading these!! Join the list!!
 
 Emily
 
 
 By bitching and bitching and bitching, they could exhaust the drama 
 of their own horror stories. Grow bored. Only then could they accept a 
 new story for their lives. Move forward.
 
 -Chuck Palahniuk, Haunted
 ___
 Histonet mailing list
 Histonet@lists.utsouthwestern.edu
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet
 
 
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] RE: Expert opinion vs. second opinion

[Gudrun Lang]

Hi,
I think the second opinion can be from your workmate next door, but the 
expert's opinion is usually from a specialist in his/her field. Usually the 
slides are sent out to another specialist facility.
Gudrun

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Sharon Scalise
Gesendet: Donnerstag, 26. Februar 2015 18:41
An: Histonet@lists.utsouthwestern.edu
Betreff: [Histonet] RE: Expert opinion vs. second opinion

I am looking for clarification about billing for expert opinion vs. second 
opinion.  Can someone explain when and how each is used and billed so that I 
can pass this along to upper management.

Thank you

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mehmet Fatih 
BOZKURT
Sent: Thursday, February 26, 2015 10:46 AM
To: Jan Shivers; Histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] CD68 and CD204 for dogs/pigs

Hello Jan,  I will stain ED1 (cd68) for you on dog tissue Friday. I hope it 
works.

On Wed, Feb 25, 2015 at 7:42 PM, Jan Shivers shive...@umn.edu wrote:

 Hi all,

 I do have MAC387 and it works great in dog and pig tissue.  The PI is 
 asking for additional CD markers to macrophages (CD68 and 204).  I 
 need them to work on dogs and pigs, but of course, there's not many 
 vendors who list species cross-reactivities on their data sheets 
 beyond the usual human and mouse.

 In my earlier trials years ago with CD68 (clone KP1), I didn't have 
 success using it on dog tissue.  Am looking for a different clone that 
 will cross-react with dogs.

 Jan

 On Wed, Feb 25, 2015 at 11:08 AM, Elizabeth Chlipala 
 l...@premierlab.com
 wrote:

  Jan
 
  MAC387 is a marker that we have used successfully in canine and 
  porcine, not entirely specific to macs but may work for your situation.
 
  LIz
 
  Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO 
  Box 18592 Boulder, CO 80308
  (303) 682-3949 office
  (303) 682-9060 fax
  (303) 881-0763 cell
  l...@premierlab.com
  www.premierlab.com
 
  March 10, 2014 is Histotechnology Professionals Day
 
  Ship to Address:
 
  Premier Laboratory, LLC
  1567 Skyway Drive, Unit E
  Longmont, CO 80504
 
  -Original Message-
  From: histonet-boun...@lists.utsouthwestern.edu [mailto:
  histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jan Shivers
  Sent: Tuesday, February 24, 2015 3:59 PM
  To: histonet
  Subject: [Histonet] CD68 and CD204 for dogs/pigs
 
  Can anyone recommend vendors for CD68 and CD204 that work on dogs 
  and
 pigs
  (on FFPE tissue)?  I don't seem to find much information on species 
  cross-reactivity online.
 
  Thanks much in advance.
 
  --
  Jan Shivers
  Senior Scientist
  IHC/Histology Section Head
  Pathology Teaching Program
  Veterinary Diagnostic Laboratory
  University of Minnesota
  1333 Gortner Ave.
  St. Paul, MN  55108
  612-624-7297
  shive...@umn.edu
  ___
  Histonet mailing list
  Histonet@lists.utsouthwestern.edu
  http://lists.utsouthwestern.edu/mailman/listinfo/histonet
 



 --
 Jan Shivers
 Senior Scientist
 IHC/Histology Section Head
 Pathology Teaching Program
 Veterinary Diagnostic Laboratory
 University of Minnesota
 1333 Gortner Ave.
 St. Paul, MN  55108
 612-624-7297
 shive...@umn.edu
 ___
 Histonet mailing list
 Histonet@lists.utsouthwestern.edu
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet




--
Asst. Prof. Dr. Mehmet Fatih BOZKURT
Department of Pathology
Faculty of Veterinary Medicine
Afyon Kocatepe University
03100, ANS Campus
Afyonkarahisar-TURKEY
Tel: +902722281312-16173/16237
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] it happened again - the foreign reply

[Gudrun Lang]

I put the sender-adress of these unwanted mails to my SPAMs. So they are
not annoying.
Gudrun

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Michelle
Gesendet: Samstag, 07. Februar 2015 16:07
An: Histonet@lists.utsouthwestern.edu
Betreff: RE: [Histonet] it happened again - the foreign reply

Interestingly enough, after I removed the histonet email address from the
quick dropdown of recently used addresses in my outlook To field and
simply manually typed the address, I did not get the foreign reply.
Related???  Who knows.

I agree with Mills, it would be nice not to see those messages.

Michelle

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Michelle
Sent: Saturday, February 7, 2015 10:01 AM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] it happened again - the foreign reply

Does anyone have a clue as to why I keep getting a foreign reply to every
email I send to histonet?  Is the group seeing the same foreign message I am
seeing?  Am I the only one getting this message?

 

Michelle

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] test

[Gudrun Lang]

test mail - best wishes to all

Gudrun

 

 

 

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] Cold plates for icing blocks?

[Gudrun Lang]

We use cold-plates. Each microtomy-place has one.  it has been working
wonderful for the last decades. After an hour switched on snow builds up
on the plate through room-moisture and as side-effect the blocks get their
moisture.
Gudrun

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Morken,
Timothy
Gesendet: Montag, 03. November 2014 20:29
An: Histonet
Betreff: [Histonet] Cold plates for icing blocks?

Does anyone use a cold plate, like that used for embedding, for icing blocks
for sectioning? Just an idea

Tim Morken
Supervisor, Histology, Electron Microscopy and Neuromuscular Special Studies
UC San Francisco Medical Center Box 1656
505 Parnassus Ave
San Francisco, CA 94143
USA

415.514-6042  (office)
tim.mor...@ucsfmedctr.orgmailto:tim.mor...@ucsfmedctr.org

CONFIDENTIALITY NOTICE: This email message, including any attachments, is
for the sole use of the intended recipient(s) and may contain confidential,
proprietary, and/or privileged information protected by law. If you are not
the intended recipient, you may not use, copy, or distribute this email
message or its attachments. If you believe you have received this email
message in error, please contact the sender by reply email and destroy all
copies of the original message.



___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] Antibody deposit visualization

[Gudrun Lang]

Hi,
 I think this is comparable with IF for detecting human autoimmun antibodies
in skin or kidney.

The human immunglobulins are detected with an fluorochrome conjugated
antibody mouse-anti-human IgG (for example).
In your case it may be successful to stain with a goat-anti-xxx IgG / IgM
/IgA etc.

On human FFPE kidney we performed pretreatment with pepsin (3-5 min) and
then 30 min polyclonal anti-serum 1:10 and 1:20.

To show immunglobulin-deposits in the glomeruli you can perform a
trichrome-stain like MSB or SFOG. Protein deposits will be shown bright red,
but are not distinguishable in the kind of protein.

It will depend on the amount of deposit, I assume.

Hope this helps
Gudrun


-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Rodriguez,
Kristofer
Gesendet: Mittwoch, 22. Oktober 2014 17:31
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Antibody deposit visualization

One of my clients has been performing experiments with a mouse model of
Dengue Virus Infection. They have been treating the mice with monoclonal
antibodies (client did not specify what antibody)as a potential treatment
for dengue virus infection. They have harvested the kidneys and put them in
10% NBF. They want to know if routine HE will show any antibody deposits.

I told him that I would not be able to directly observe the antibody deposit
itself, however I might be able to observe the reaction to the antibody
deposit in the basement membrane of the glomeruli as a membranous
glomerulopathy on an HE.

I was wondering if I should do any special stains that might show more than
an HE.
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] Neutrophil staining

[Gudrun Lang]

What about PAS staining? Granulozyte granula are PAS positiv.
Gudrun

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Connolly, 
Brett M
Gesendet: Montag, 08. September 2014 15:30
An: Hans B Snyder; histonet@lists.utsouthwestern.edu
Betreff: RE: [Histonet] Neutrophil staining

Hello Hans,

We had very good staining using Ly-6B.2 from Serotec (MCA771)  on C57B6 mice, 
but needed to switch to Ly-6G for CH3 mice.  The MCA771 datasheet tells you 
which strains react with that antibody.

Brett

Brett M. Connolly, Ph.D.
Principle Scientist, Imaging Dept.
Merck  Co., Inc.
PO Box 4, WP-44K
West Point, PA 19486
brett_conno...@merck.com
T- 215-652-2501
F- 215-993-6803



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Hans B Snyder
Sent: Saturday, September 06, 2014 5:43 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Neutrophil staining

Hello All,

I am looking to  stain neutrophils using special stains and antibodies in mice 
tissue.  I wanted your opinion about the best methods to do so.  I am thinking 
of using the may Grunwald giemsa and T-blue for specials and myeloperoxidase 
and Ly6G for the antibodies.

What are your favorite special stains and antibodies to stain for neutrophils?

Best regards

Hans B Snyder
Histologistics
60 Prescott Street
Worcester, MA 01605
508-308-7800
h...@histologistics.com

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Notice:  This e-mail message, together with any attachments, contains 
information of Merck  Co., Inc. (One Merck Drive, Whitehouse Station, New 
Jersey, USA 08889), and/or its affiliates Direct contact information for 
affiliates is available at
http://www.merck.com/contact/contacts.html) that may be confidential, 
proprietary copyrighted and/or legally privileged. It is intended solely for 
the use of the individual or entity named on this message. If you are not the 
intended recipient, and have received this message in error, please notify us 
immediately by reply e-mail and then delete it from your system.


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] RE: whole slide scanners

[Gudrun Lang]

Hi,
for which purpose do you use the high troughput scanner. Archive or
diagnostics?
Gudrun

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Elizabeth
Chlipala
Gesendet: Dienstag, 05. August 2014 17:16
An: James Watson; 'Kalleberg, Kristopher';
histonet-requ...@lists.utsouthwestern.edu; histonet@lists.utsouthwestern.edu
Betreff: [Histonet] RE: whole slide scanners

I agree with James the type of scanner you need will be dependent upon your
use cases and workload.  We have an Aperio ScanScope XT which is an 120
slide scanner and that works well for us.

I'm going to make a shameless plug for a workshop that myself, Bill DeSalvo
and Jesus Ellin will be giving at the annual meeting in Austin this year,
it's called Digital Pathology for the Histotech - A Guide to Implementation
it's an all day workshop that is on Saturday the 23rd and it will cover all
aspects of how to select a scanner, how to implement scanning, etc.  It will
be very valuable to histotechs that are already scanning and also techs that
are new to the scanning process.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box
18592 Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
l...@premierlab.com
www.premierlab.com

March 10, 2014 is Histotechnology Professionals Day

Ship to Address:

Premier Laboratory, LLC
1567 Skyway Drive, Unit E
Longmont, CO 80504

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of James Watson
Sent: Tuesday, August 05, 2014 8:14 AM
To: 'Kalleberg, Kristopher'; histonet-requ...@lists.utsouthwestern.edu;
histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: whole slide scanners

The scanner you get depends on your usage and volume.  Three that I
recommend are for high volume.

Aperio, 
Good image quality,  if you need to link to study metadata Spectrum software
is great.  Recommend for chromogen. Separate scanner for fluorescence,
fluorescent scanner  only holds 5 slides at a time.  

Hamamatsu Nanozoomer,   
What we use,  we do high volume chromogen and fluorescence,  excellent for
both.  Our NanoZoomer holds 210 slides for a chromogen run.  Can load up
about 100 fluorescent slides per night,  auto focus works well.  

Philips,
Chromogen only,  very fast scanning with high quality scans.  60 slides per
hour.

There are many others on the market for smaller workloads.

James Watson HT  ASCP
GNF  Genomics Institute of the Novartis Research Foundation Scientific
Technical Leader II, Histology Tel    858-332-4647 Fax   858-812-1915
jwat...@gnf.org


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kalleberg,
Kristopher
Sent: Tuesday, August 05, 2014 5:18 AM
To: histonet-requ...@lists.utsouthwestern.edu;
histonet@lists.utsouthwestern.edu
Subject: [Histonet] whole slide scanners

Hello All,

I am looking into the purchase of a whole slide scanner.  If anyone could
supply some recommendations it would be greatly appreciated.  Thank you.

Kris
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] RE: whole slide scanners

[Gudrun Lang]

Does anyone out there use digital pathology for routine diagnostics? 
Gudrun

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Gudrun
Lang
Gesendet: Dienstag, 05. August 2014 19:13
An: 'Elizabeth Chlipala'
Cc: histonet@lists.utsouthwestern.edu
Betreff: AW: [Histonet] RE: whole slide scanners

Hi,
for which purpose do you use the high troughput scanner. Archive or
diagnostics?
Gudrun

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Elizabeth
Chlipala
Gesendet: Dienstag, 05. August 2014 17:16
An: James Watson; 'Kalleberg, Kristopher';
histonet-requ...@lists.utsouthwestern.edu; histonet@lists.utsouthwestern.edu
Betreff: [Histonet] RE: whole slide scanners

I agree with James the type of scanner you need will be dependent upon your
use cases and workload.  We have an Aperio ScanScope XT which is an 120
slide scanner and that works well for us.

I'm going to make a shameless plug for a workshop that myself, Bill DeSalvo
and Jesus Ellin will be giving at the annual meeting in Austin this year,
it's called Digital Pathology for the Histotech - A Guide to Implementation
it's an all day workshop that is on Saturday the 23rd and it will cover all
aspects of how to select a scanner, how to implement scanning, etc.  It will
be very valuable to histotechs that are already scanning and also techs that
are new to the scanning process.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box
18592 Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
l...@premierlab.com
www.premierlab.com

March 10, 2014 is Histotechnology Professionals Day

Ship to Address:

Premier Laboratory, LLC
1567 Skyway Drive, Unit E
Longmont, CO 80504

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of James Watson
Sent: Tuesday, August 05, 2014 8:14 AM
To: 'Kalleberg, Kristopher'; histonet-requ...@lists.utsouthwestern.edu;
histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: whole slide scanners

The scanner you get depends on your usage and volume.  Three that I
recommend are for high volume.

Aperio, 
Good image quality,  if you need to link to study metadata Spectrum software
is great.  Recommend for chromogen. Separate scanner for fluorescence,
fluorescent scanner  only holds 5 slides at a time.  

Hamamatsu Nanozoomer,   
What we use,  we do high volume chromogen and fluorescence,  excellent for
both.  Our NanoZoomer holds 210 slides for a chromogen run.  Can load up
about 100 fluorescent slides per night,  auto focus works well.  

Philips,
Chromogen only,  very fast scanning with high quality scans.  60 slides per
hour.

There are many others on the market for smaller workloads.

James Watson HT  ASCP
GNF  Genomics Institute of the Novartis Research Foundation Scientific
Technical Leader II, Histology Tel    858-332-4647 Fax   858-812-1915
jwat...@gnf.org


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kalleberg,
Kristopher
Sent: Tuesday, August 05, 2014 5:18 AM
To: histonet-requ...@lists.utsouthwestern.edu;
histonet@lists.utsouthwestern.edu
Subject: [Histonet] whole slide scanners

Hello All,

I am looking into the purchase of a whole slide scanner.  If anyone could
supply some recommendations it would be greatly appreciated.  Thank you.

Kris
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] Fwd: SOP on embedding tissues of different sizes in same mold

[Gudrun Lang]

We avoid to put specimens of different size into one cassette in general
(especially with biopsies). If I have too pieces of different size with a
clear cut side, I put them both on the ground of the mold. I think one could
never precisly put a specimen in a certain level into the mold, if it
doesn't lie on the bottom.
Gudrun

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Sanjeet
Gesendet: Donnerstag, 31. Juli 2014 03:44
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Fwd: SOP on embedding tissues of different sizes in same
mold



Sent from my iPhone

Begin forwarded message:

 From: Sanjeet asanj...@yahoo.com
 Date: July 30, 2014 at 9:15:25 PM EDT
 To: histonet-requ...@lists.utsouthwestern.edu
histonet-requ...@lists.utsouthwestern.edu
 Subject: SOP on embedding tissues of different sizes in same mold
 
 Hi
 Does anyone in the Histo world have an SOP on how to embed tissues of
different sizes in the same mold. Do you embed in different levels depending
upon the size, the larger embedded first and the smallest at the end. Or do
you embed all in the same plane regardless of the tissue size. Are you able
to justify and salvage all tissue on the slide.
 Thank you all in advance
 Sanjeet 
 
 
 Sent from my iPhone
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] Fwd: Embedding skin

[Gudrun Lang]

As a first information, we use sliding microtomes with the knife set in an
angel (called declination).
We embed straight, because areas of same concistancy should be positioned in
one line and in the cutting direction.
Areas of more rigidity compress the smoother ones, if they are shifted in an
angle. 

If you have problems with the orientation of the tissue in the block, just
turn the blockholder.
Gudrun



-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Sanjeet
Gesendet: Donnerstag, 31. Juli 2014 03:46
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Fwd: Embedding skin



Sent from my iPhone

Begin forwarded message:

 From: Sanjeet asanj...@yahoo.com
 Date: July 30, 2014 at 9:27:46 PM EDT
 To: histonet-requ...@lists.utsouthwestern.edu
histonet-requ...@lists.utsouthwestern.edu
 Subject: Embedding skin
 
 Hi Histo techs
 
 Need some info on the correct orientation of skin tissue. Punch/ ellipse.
Does anyone have a literature on this topic. I am used to embed large skin
in an angle, the skin being on top. 
 Currently the place where I work have different protocol, the skin is
embedded straight, the epidermis being right angled to the mold.
 I find the section difficult to cut when the skin is embedded straight ,
the section are compressed and more chances scoring along the section.
 
 Thanks 
 Sanjeet
 
 Sent from my iPhone
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] problems Sllde Mate print

[Gudrun Lang]

Dear histonetters,

we have problems with the stability of the print of our slide mates
(Thermo).

We use regular slides with frosted end. The print usually looks good and
stable at the time of printing.

But after staining the print comes off like peeled skin. Very easy to
remove.

 

We take care, that the print doesn't come in contact with organic solvents
(deparaffination reagens = Histo SAV, butyl acetate), but without any
logical reason the first line of the print peels off and the second and
third line is rather ok.

 

Has anyone seen similar problems and solved them?

 

Gudrun Lang

 

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] Ideas for a Retiring Pathologist

[Gudrun Lang]

Buy him a ticket to Austria and let him work here with us. ; )  . still
looking for pathologists

Gudrun

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von
lori.dis...@hcahealthcare.com
Gesendet: Dienstag, 01. Juli 2014 17:25
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Ideas for a Retiring Pathologist

I'm looking for creative ideas to celebrate a retiring pathologist sometime
this year.  What have you all done in the past?

Lori Disher


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Pathologists?

[Gudrun Lang]

Dear histonetters!

Maybe you can help me.  We are looking for pathologists for our clinical
histolab in Linz, Austria. Anyone (German speaking), who is interested in is
welcome to inform him- or herself on this website.

http://www.linz.at/akh/10374.asp

 

We have a well equipted, modern histolab (from routine histo till FISH and
molecularpathology). 

Best regards

Gudrun 

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] Decal and molecular testing

[Gudrun Lang]

Hi,
we had success with 5% formic acid on trephine bone marrow for  DNA-PCR
(BRAF, c-kit mutation) and RNA-ISH (kappa), but had no success for
FISH-testing.
Perhaps someone can recommend a method, that also supports FISH?
Gudrun Lang

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von O'Donnell,
Bill
Gesendet: Dienstag, 11. März 2014 19:25
An: 'histonet@lists.utsouthwestern.edu'
Betreff: [Histonet] Decal and molecular testing

Can anyone recommend a decal solution that does no damage to specimens for
molecular testing - or one that has minimal damage? Thanks - Bill


William (Bill) O'Donnell, HT (ASCP) QIHC Senior Histologist Good Samaritan
Hospital
10 East 31st Street
Kearney, NE 68847 

SERENITY is not freedom from the storm, but peace amid the storm.

Cultivate it in PRAYER!

 


 

This electronic mail and any attached documents are intended solely for the
named addressee(s) and contain confidential information. If you are not an
addressee, or responsible for delivering this email to an addressee, you
have received this email in error and are notified that reading, copying, or
disclosing this email is prohibited. If you received this email in error,
immediately reply to the sender and delete the message completely from your
computer system.

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] microwave processing

[Gudrun Lang]

Thank you for the kind responses.
Gudrun

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Gudrun
Lang
Gesendet: Montag, 13. Jänner 2014 19:56
An: Histonet@lists.utsouthwestern.edu
Betreff: [Histonet] microwave processing

 

Hi!

Can someone recommend literature about microwave processing. I'm interested
in the physical principles behind the process. And I want to get answers to
the questions: why is this microwave-assisted infiltration faster? What
happens to proteins /antigens under microwave radiation? Is there a
difference between conventional or microwave processing in relation to
antigen preservation after usual formalinfixation.

 

Thanks in advance

Gudrun Lang

 

 

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] microwave processing

[Gudrun Lang]

 

Hi!

Can someone recommend literature about microwave processing. I'm interested
in the physical principles behind the process. And I want to get answers to
the questions: why is this microwave-assisted infiltration faster? What
happens to proteins /antigens under microwave radiation? Is there a
difference between conventional or microwave processing in relation to
antigen preservation after usual formalinfixation.

 

Thanks in advance

Gudrun Lang

 

 

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] AW:cont. workflow

[Gudrun Lang]

Hi Peggy!
You talk about a lab with many specimens, many blocks.
What do you think is the number of blocks, where a change from batch to
flow makes sense?

We have about 300 blocks per day and work from 6.30 till 15.30. Usually we
are ready with slide-distribution at noon. And we do also IHC, FISH,
mutation analysis, IF , many fast frozens- so we are busy, when we are not
dealing with routine-histology.
And we are happy with our system. ;-) Fixation is all in a comparable range.
TAT of biopsies is one day after entry (patients are called in usually one
week afterwards from the clinicians...). And we have a very diversified
workplace with comfortable workshifts.

Advisers often hold up cont. workflow as best, but I think it must fit to
the circumstances!

Gudrun Lang


-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Lee 
Peggy Wenk
Gesendet: Montag, 06. Jänner 2014 18:31
An: Podawiltz, Thomas; Deanna Leslie; histonet@lists.utsouthwestern.edu
Betreff: Re: [Histonet] Soaking artifact

In most labs, the processor runs all night long. Someone comes in very early
in the morning, empties the processor, starts the purge cycle, and then
starts embedding a lot of blocks.

The tissue processor then sits, doing nothing, from after the purge in the
early morning, until sometime in the late afternoon, when all the tissues
are loaded in for the overnight run. That means the tissue processor is
doing nothing for up to 12 hours during the daytime.

How about, besides the overnight run, we can set up 1 or 2 other shorter
runs during the day, with the small biopsies.

How about - process all the large tissues overnight, but keep the little
biopsies that you grossed all afternoon in formalin until the morning. Empty
out the large tissues, purge the process, embed the large tissues and start
microtoming them.

After the purge is done, put the small biopsies from the afternoon on the
tissue processor, and process them for 1.5-2 hours (and if your processor is
able to process half a load, do that to save on reagents). Then embed them
and start microtoming them. Purge the processor again.

In the mean time, all morning, collect the small biopsies again. After
lunch, short process all the small biopsies from the morning. Embed them in
the afternoon, purge the processor again, and load up the overnight load.

If you don't have time to microtome the morning small biopsies (that you
embedded in the afternoon), someone can microtome them in the morning the
next day. Either with the large overnight load, or have someone else come in
early, and while the other people are embedding the large tissue overnight
load, they can be microtoming the small biopsies that were embedded in
previous afternoon.

Yes, all of this will mean changes:
- staggered hours that people will be coming in
- processing, embedding and microtoming continuously throughout the day
- someone might have to microtome more than someone else, or might have to
embed more than someone else. But if you rotate jobs around, over the
months, everyone ends up doing the same amount of work overall.

This is a type of continuous work flow, and does lead to faster turn around
time and efficiency.

When our lab changed to this system (we are an 1100 bed hospital, with lots
of tissues from our ORs, from outside hospitals and clinics and doctors
offices, so lots and lots of blocks), it took getting everyone involved -
people accessioning, grossing, the histotechs, and the pathologists (they
were not going to get their slides in numerical order). We have short
cycles, the overnight long cycles, some rush cycles, and long cycles for
breast and autopsy brain. We actually have more than 3 runs, but then we are
working almost 24/7. During the time we were switching to continuous work
flow, we had a few histotechs off on maternity and/or medical leaves. And we
got a couple more clients, so the work load increased. But because of the
continuous work flow, we were able to handle the additional work without
having to hire anyone.

Whereas before, we would process ALL the blocks overnight, and then would
have lots of people embedding lots of blocks first thing in the very early
morning, and then having to put in the in order, and no one could start
microtoming until all the blocks were embedding and in order (so some days
the microtoming techs were sitting around with nothing to do for a time). 
Then, everyone had piles of blocks to cut, for blocks 100-150 were going to
be cut hours and hours after everyone started microtoming blocks 1-50. Then,
there were racks and racks of slide piling up to be stained with HE (and at
that point, we were still labeling after staining). So there were all these
spots where tissue was being held up, unnecessarily:
- waiting to be processed
- waiting to be embedding
- waiting to be microtomed
- waiting to be stained
- waiting to be labeled
- then the pathologists

AW: [Histonet] Bone samples long-term storage in 10% formalin or 4% paraformaldehyde

[Gudrun Lang]

Paraformaldehyd is formaldehyd in solid form. Formalin is the aequous
solution of formaldehyd. 
So the main characteristics are the same.

Gudrun Lang

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Orla M
Gallagher
Gesendet: Donnerstag, 05. Dezember 2013 19:31
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Bone samples long-term storage in 10% formalin or 4%
paraformaldehyde

Dear Histonetters,

What is your opinion on storing bone samples long-term (more than a couple
of weeks) in 10% formalin? As I was taught, best practice has always been to
fix only as long as necessary, depending on the size of the sample, then
decalcify and process to wax, and I always stress this to everyone I advise.

However, research colleagues sometimes wish to do histology on bone samples
that have been stored for months ..or even years! As the formalin pH becomes
more acidic, there is formalin pigment and the immunoreactivity and TRAP
enzyme activity is diminished or destroyed during long fixation, is there
any way of minimising this e.g. has anyone tried regularly replacing the old
formalin with fresh buffered formalin, or storing formalin-fixed bones in
any other medium? I'm also interested in how best to fix in 4%
paraformaldehyde and whether the problems are the same with long-term
storage.

Thanks for your comments.

All the best,
Orla

--
**
Ms. Orla Gallagher
Bone Analysis Laboratory
Mellanby Centre for Bone Research
Department of Human Metabolism
D Floor Medical School
University of Sheffield
Beech Hill Road
Sheffield
S10 2RX
UK

Website: http://mellanbycentre.dept.shef.ac.uk

Tel: 0044114-2713337 (office)
  0044114-2713174 (lab)
E-Mail:o.m.gallag...@sheffield.ac.uk


*STOP*: Do you really need to print this e-mail?

*BE GREEN:* Keep it on the screen.


*Times Higher Education University of the Year*



Data protection and confidentiality:
The information contained in this message or any appended documents may be
privileged and confidential and is intended for the exclusive use of the
addressee(s). If you are not the addressee, any disclosure, reproduction,
distributions, other dissemination or use of this message is strictly
prohibited and may be unlawful. If you receive this correspondence in error
please contact the sender immediately and permanently delete/destroy what
you have received.
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] Over fixation

[Gudrun Lang]

Formaldehyd binds as methylol-group to basic functional groups on proteins.
The longer the fixation the longer the possibility to bind. The same basic
groups bind acid dyes like eosin. 
On the other side leads acidic formalin to degradation of DNA. This can lead
to lower affinity of hematoxylin.
So in sum long fixed tissue should stain paler. In old literature you find
the recommendation to wash fixed tissue in tapwater as long as the fixation
has longed. Hmm ;-)
Gudrun

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Sanjeet
Gesendet: Sonntag, 24. November 2013 21:33
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Over fixation

Hi Histo folks,
Tissue biopsy that are  not fixed properly do have an adverse effect on H 
E staining. Tissues can be kept in formalin for a period of 5 to 7 days as
per literature. But what happens to the tissues that are kept over for a
longer period of time. By that I mean 6 to 12 months. Would over fixed
tissues have any effect on H  E stain. Some of my colleagues say NO, but I
say YES. I find that the over fixed tissues lose their affinity towards the
dyes . Please advice. 

Sent from my iPhone
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] Rolling of my ribbon ? Another paraffin question.

[Gudrun Lang]

Has this new paraffin a higher melting point than the former? It may happen
that the block/paraffin is too hard at your cutting temperature to stick.
Does it make ribbons after a while trying?
Gudrun

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Stella
Mireles
Gesendet: Mittwoch, 13. November 2013 21:18
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Rolling of my ribbon ? Another paraffin question.

*I am presently using a paraffin designated as an IM product.* *We are a
facility that cuts only autopsies and have been experiencing alot of rolling
of our sections.* *We did recently switch to this product, because of cost.*

*Question :  Is the paraffin you are using working well on autopsy tissue
and producing ribbons right away ? Do you use it for infiltration as well as
embedding ?*

*Thank you for your assistant.*


*Stella Walters*
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] fixation question

[Gudrun Lang]

Hi!
In my opinion 24 hours should be the minimal duration for fixation in 4%
buffered formaldehyde. The shorter the time the more differences may occur.
It is important to prepare the specimens in the same way at the grossing -
means same size of tissue in the cassette or at least minimal size of 3 mm
diameter.  24 hours of a whole organ in formalin is not the same as 24 hours
of a small tissue-block.
Look at the article of Cecil Fox about formaldehyde. He says that shrinkage
is not due to formalin, but more caused by the processing, when lipids and
water are removed.
The better the crosslinking, the less sensitive is the tissue to shrinkage
or swelling.
Immunhisto can be adopted to longer fixation, but too short fixation may
give impact on the morphology, that can't be recovered.

Shrinkage during processing can be intensified by very long times in
absolute ethanol and xylen. Let you inform about the processing protocol of
the company. The protocol should fit to your specimens.
On the other side inform the company about your fixation times. They may
prolong the antigen retrieval for better results.
My advice without knowledge of your special experiment is a fixationtime of
24-72 hours. 

Bye
Gudrun Lang


-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von
ncose...@siumed.edu
Gesendet: Samstag, 19. Oktober 2013 16:51
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] fixation question

Hello Histonetters:

Our lab has a resident wanting to start a project involving drug effect on
tumor size.  Since he is wanting to compare tumor size at certain time
points after drug treatment (and do IHC staining intensity), he wonders how
much affect varying fixation times will have on the tissue.  We are not
doing the processing or embedding ourselves so we can't control for such
variation.

Is this a legitimate concern? If one piece of tissue sits in fix 24 hours
before processing/embedding, will the morphology be drastically different
than a second piece that sits longer (or shorter) than 24 hours?

Is so, would fixing the tissue ourselves for a set time and then putting in
some sort of buffer or ethanol before we ship the tissue to the company who
embeds it make a difference? The resident is really trying to minimize
swelling or excess shrinkage of the tissue.

Thanks!


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] HT HistoDeck question...

[Gudrun Lang]

We use 36-40% formaldehyde for a minimal time of 2-3 dips for fast frozen
HE. On the other side the slides stand in the solution as long as all
frozens are already cut.
Commercial formaldehyde contents a good part of methanol (a MSDS says
5-15%). So fast fixation is a combination of formaldehyde and alcohol
fixation.
Additional we use Harris hematoxylin, that's also mainly made of ethanol. 
So in my opinion all together fixation is rather due to alcohol in this way.


Gudrun 
 

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Rathborne,
Toni
Gesendet: Donnerstag, 03. Oktober 2013 20:34
An: 'Jennifer MacDonald'; Grantham, Andrea L - (algranth)
Cc: HISTONET; histonet-boun...@lists.utsouthwestern.edu
Betreff: RE: [Histonet] HT HistoDeck question...

For those of you who use the 40% formaldehyde, how long is you fixation time
on frozen slides? We use 10% NBF with 1 minute to fix, but sometimes it gets
hectic if you have multiple frozens all at once. 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jennifer
MacDonald
Sent: Thursday, October 03, 2013 2:04 PM
To: Grantham, Andrea L - (algranth)
Cc: HISTONET; histonet-boun...@lists.utsouthwestern.edu
Subject: Re: [Histonet] HT HistoDeck question...

We also use this oil red O method and use the 40% formaldehyde.
The questions lacks enough information to correctly answer it.  I am sure
the author of the question had something in mind and other options didn't
occur to him/her at the time.
Jennifer




From:   Grantham, Andrea L - (algranth) algra...@email.arizona.edu
To: 
Cc: HISTONET histonet@lists.utsouthwestern.edu
Date:   10/03/2013 08:33 AM
Subject:Re: [Histonet] HT HistoDeck question...
Sent by:histonet-boun...@lists.utsouthwestern.edu



I'd go with A, but it really depends on what you are going to do with the
sections after fixation.

In the protocol for Oil Red O in Freida's second edition (that I use almost
daily combined with some steps from PolyScientific's ORO protocol), step #1
says to fix in 40% formaldehyde. Doesn't say vapors so I plunk the slides in
the liquid 40%.
I have fixed frozen sections for regular HE and other stains as well in 40%
formaldehyde and they come out beautiful.
Unless there is some specific request or reason to use cold acetone or
alcohol this is what I use.

Andrea Grantham, HT (ASCP)
Senior Research Specialist
University of Arizona
Cellular and Molecular Medicine
Histology Service Laboratory
P.O.Box 245044
Tucson, AZ 85724

algra...@email.arizona.edumailto:algra...@email.arizona.edu
Tel: 520.626.4415 Fax: 520.626.2097





___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Benchmark Ultra troubles

[Gudrun Lang]

Hi Ventana-users,

we have a special technical problem with our Ultra. It seems, that the
liquid in the waste-tube is sometime pressed out of the hole on the platform
like a fountain.

You can imagine the nice surprise, when the oil drops from the Ultra-roof
onto the carousel-top. Everything is oily.

The technical service has'nt found a cause until now. The filter in the tube
has been changed.

Has anyone seen a similar problem? Solved the problem?

 

Gudrun Lang

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] IHC trun-around-time

[Gudrun Lang]

Hi Tom,
we also do same-day-IHC. Our cut off is 10.30. We use Benchmark ultra with
ultravision-kit. The run is started between 10.30-10.50 and needs about 3
hours (more or less). After the run is completed we need about 30 hours
until delivering the slides.

For faster processing we have an online order service for IHC. The doctors
fill a Word-form and print it on our lab-printer. So the orders go in
continously.
The urgent cases (mamma-biopsies eg.) are cut in advance with the HE and
relatively fast prepared for IHC.
Usually the slides need no drying time in the oven.

What issues? What changes?

Regards
Gudrun Lang


-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Tom
McNemar
Gesendet: Dienstag, 01. Oktober 2013 17:00
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] IHC trun-around-time

Hello all,

I am wondering how many places offer same-day staining of IHC.  We are a
small hospital based lab that averages about 70 IHC slides per week.  We
operate 5 days/week, 0600-1600.  We give same day staining for IHC requests
that are received by 10am and are using the Ventana Benchmark XT.   Due to
some issues that have developed, we may have to change the way we do things.

With our current 10am cutoff plus possible pre-analytical changes the
instrument will finish too late in the afternoon.  The pathologists really
like the same day service on IHC but I'm thinking that I may have to make
the cutoff time earlier and anything after that will just have to wait.

So I am wondering if anyone else of similar size, instrumentation, and
workload, offers same-day staining for IHC.  I appreciate your input.
Thanks.


Tom McNemar, HT(ASCP)
Histology Co-ordinator
Licking Memorial Health Systems
(740) 348-4163
(740) 348-4166
tmcne...@lmhealth.orgmailto:tmcne...@lmhealth.org
www.LMHealth.orgfile:///C:\Documents%20and%20Settings\TMCNEMAR\Application%
20Data\Microsoft\Signatures\www.LMHealth.org


This e-mail, including attachments, is intended for the sole use of the
individual and/or entity to whom it is addressed, and contains information
from Licking Memorial Health Systems which is confidential or privileged. If
you are not the intended recipient, nor authorized to receive for the
intended recipient, be aware that any disclosure, copying, distribution or
use of the contents of this e-mail and attachments is prohibited. If you
have received this in error, please advise the sender by reply e-mail and
delete the message immediately. You may also contact the LMH Process
Improvement Center at 740-348-4641. E-mail transmissions cannot be
guaranteed to be secure or error-free as information could be intercepted,
corrupted, lost, destroyed, arrive late or incomplete, or contain viruses.
The sender therefore does not accept liability for any errors or omissions
in the contents of this message, which arise as a result of e-mail
transmission. Thank you.
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] edge effect IHC

[Gudrun Lang]

 

Hi!

Can someone give me a nice description of the edge effect in IHC? Is there
a common opinion about the causes?

 

Thank you

Gudrun Lang

 

 

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] Re: edge effect IHC

[Gudrun Lang]

Thank you for your kind responses.
Are there any circumstances when an edge-effect could be misinterpreted as a
specific nuclear staining like ER-IHC?  =false positive staining

Gudrun

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Mehmet
Fatih BOZKURT
Gesendet: Mittwoch, 11. September 2013 19:05
An: Teri Johnson; Histonet@lists.utsouthwestern.edu
Betreff: Re: [Histonet] Re: edge effect IHC

Sometimes edges of tissues can get more stain due to disruption of edge's
cells more than center during the antigen retrieval. But this is not  false
positivity. It must pay attention to this situation


On Wed, Sep 11, 2013 at 7:50 PM, Teri Johnson tjohn...@gnf.org wrote:

 Ditto to what Tim said (as usual!) What is often referred to as edge 
 artifact is sometimes excellent staining due to the edges fixed early 
 and well, and the interior of the tissue poorly penetrated and not as 
 well fixed. You can usually tell because the staining pattern is 
 correct and well demarcated, and easily distinguishable from the type 
 seen in drying artifact.

 Teri Johnson
 Manager, Histology
 Genomics Institute for
 Novartis Research
 Foundation
 San Diego, CA
 858-332-4752

 ___
 Histonet mailing list
 Histonet@lists.utsouthwestern.edu
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet




--
Mehmet Fatih BOZKURT, DVM, PhD
Afyon Kocatepe University
Faculty of Veterinary Medicine
Department of Pathology
03030, ANS Campus
Afyonkarahisar-TURKEY
Tel: +902722281312-16173/16237
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] FISH enumeration

[Gudrun Lang]

Pathologists do FISH reading. Learning by doing. Manual enumeration.

Each doctor, who orders a FISH has to read it by him/herself. Often they do
it with a second person/second look for reliable results.

Gudrun Lang
Histolab,  Linz, Austria


-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von joelle
weaver
Gesendet: Samstag, 07. September 2013 16:22
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] FISH enumeration

 For any laboratories out there who perform in house FISH procedures, if you
could share what personnel are responsible for doing the signal enumeration
 scoring? It would be helpful if you could describe the personnel's
training and certification,  as well as an approximation of FTE's needed
with some volumes. Do you do manual enumeration or use scanning software? 
Thanks for any input.





Joelle Weaver MAOM, HTL (ASCP) QIHC
 
 From: tmcne...@lmhealth.org
 To: thisis...@aol.com; histonet@lists.utsouthwestern.edu
 Date: Fri, 6 Sep 2013 05:45:41 -0400
 Subject: RE: [Histonet] Cell Block Preparation
 CC: 
 
 This is how we do it now.  In the old days, we used agar and to my mind,
it is still the best way when you have scant material.
 - Spin in a conical tube and pour off
 - Melt an agar slant (we get TSA slant from micro)
 - Pour the agar into the conical tube and spin for 5 minutes
 - The agar will re-solidify and whatever sediment there is will be
concentrated in the very tip of the cone
 - The agar will slide out of the centrifuge tube
 - Slice off the very tip and wrap in lens paper
 - Place the wrapped tip in a cassette and process as usual
 - Embed the specimen tip down and you are good to go...
 
 I still use this method today when I feel it necessary.  Works great.
 
 Tom McNemar, HT(ASCP)
 Histology Co-ordinator
 Licking Memorial Health Systems
 (740) 348-4163
 (740) 348-4166
 tmcne...@lmhealth.org
 www.LMHealth.org
 
 
 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Ann Specian
 Sent: Thursday, September 05, 2013 12:45 PM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] Cell Block Preparation
 
 
 I am getting complaints in regard to insufficient cell blocks.  We
currently spin, pour off the supernatant, retrieve the sediment and process
in lens paper.
 
 Does anyone have a more current technique which renders better
cellularity?
 
 Also, do you know which renders a better cell block:  a fresh specimen, a
specimen fixed in Cytolyt or a specimen fixed in 10% NBF?
 
 Thanks,
 Ann
 ___
 Histonet mailing list
 Histonet@lists.utsouthwestern.edu
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet
 
 This e-mail, including attachments, is intended for the sole use of the
individual and/or entity to whom it is addressed, and contains information
from Licking Memorial Health Systems which is confidential or privileged. If
you are not the intended recipient, nor authorized to receive for the
intended recipient, be aware that any disclosure, copying, distribution or
use of the contents of this e-mail and attachments is prohibited. If you
have received this in error, please advise the sender by reply e-mail and
delete the message immediately. You may also contact the LMH Process
Improvement Center at 740-348-4641. E-mail transmissions cannot be
guaranteed to be secure or error-free as information could be intercepted,
corrupted, lost, destroyed, arrive late or incomplete, or contain viruses.
The sender therefore does not accept liability for any errors or omissions
in the contents of this message, which arise as a result of e-mail
transmission. Thank you.
 
 ___
 Histonet mailing list
 Histonet@lists.utsouthwestern.edu
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet
 
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] microwave processing plus embedding

[Gudrun Lang]

Hi!

Has anyone heard about a new device for automatic microwave processing and
embedding (not the Sakura Xpress) ?

It should be a two-bath processor and should use Isopropanol.

 

For about 45 samples per run.

 

Thanks for your responses in advance

 

Gudrun Lang

Ltd. BMA histolab Linz, Austria

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] Bone/cartilage/epithelial tissue stain

[Gudrun Lang]

Rui,
in your first post about your problem, you wrote about decalcification of
your samples. 
Are these samples, where you want to demonstrate bone, decalcified? Which
decalcifier?
Haematoxylin stains only non-decalcified bone, due to the Ca-ions.
Strong acids for decalcification (like HNO3 or HCl) may alter the
tissue-stainability. They are said to solve some amount of proteoglycans.
If they are not decalcified, you can combine a von-Kossa-stain with a
trichrome-stain or alcianblue. Calcified bone stains black.

An easy stain would be the combined Alcianblue-PAS. Alcianblue demonstrates
acid proteoglycans, PAS demonstrates neutral glycoproteins like collagen.
But I think, proteoglycans and collagen are in such a tight junction in
cardilage, that it will give a mixed dye-appearance. But worth a try.

Gudrun

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Rui TAHARA
Gesendet: Freitag, 23. August 2013 09:49
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Bone/cartilage/epithelial tissue stain



Hi,
Would anyone suggest me what staining is best to color differentiate between
cartilage and bone and epithelial tissues in avian embryos? 

I have been trying Mallory Trichrome for embryos but recently I was
suggested that Mallory Trichrome stains cartilage differently in embryos
compared to adult samples since Aniline blue stains fiber that may not
develop in early embryos. There is some protocol that modified the Mallory
Trichrome that could be applied to embryos. However, the resulting colors of
all tissues look all purple-ish and difficult to tell the cartilage from the
weak blue stain from aniline blue. 

Currently I am thinking to try out Alcian blue/Hematoxylin and Eosin stain
(Ehrlich’s hematoxylin). The purpose of the staining is to look at
interaction between ossification and epithelial development so I think
alcian blue for staining cartilage works but I am wondering if there is any
other staining combination with alcian blue exist for visualizing bone and
epithelial tissue (e,g. alcian blue/alizarine red with other staining?). 

 

Any suggestion would be appreciated! 

Rui TAHARA
PhD Candidate
Biology Department
McGill University




 

  


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] Uncertified Histotechs

[Gudrun Lang]

It's interesting to see the diference in aquisition of a person for
histolab. Here in Austria only biomedical scientist have the legal right for
working with clinical samples. Therefore one part of education deals also
with histotechnology and they have a few practical hours on microtome.
But I would never expect a newborn BMA to be able to cut from the very
beginning. After a few weeks in the grossing room, new BMAs learn real
cutting on clinical samples and they are given three to four weeks at least
to learn the whole spectrum (from appendix to core-biopsies).
It has some advantages to take the own hands on the new coworker. (Besides,
as far as I remember, we never got a new histotech from another histolab in
the last 20 years.)
On the other hand new employees can be easily signed off in the first 6
months, if they don't reach the trainings-aims. 
So I think, skilled people learn cutting fast enough. More important, from
my point of view, is,  that they have already learned the theoretical
background. Because theory-learning beside working (especially at home) is
often assumed as impossible.

Gudrun




-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Sullivan,
Beatrice
Gesendet: Freitag, 23. August 2013 14:29
An: joelle weaver; Jon Hannasch; histonet@lists.utsouthwestern.edu
Betreff: RE: [Histonet] Uncertified Histotechs

While I understand the need for certification and continuing education,
because of regulations it is very hard to even have your OJT's sit for their
certification. Recently I interviewed candidates for an open position at my
facility. One candidate in particular looked very good on paper. I brought
this person in for an interview. Candidate was certified by CAP as a Histo-
Technician. I always test their microtomy skills. Imagine my dismay when
after more than I hour this person had not produced one decent slide.
Needless to say the interview was pretty much over. Lesson here is that
letters after one's  name does not make a good Histo-Tech.

Beatrice Sullivan HT(ASCP)HTL  CLSP(NCA) Corporate Histology Supervisor
Virtua,Voorhees

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of joelle
weaver
Sent: Friday, August 23, 2013 8:00 AM
To: Jon Hannasch; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Uncertified Histotechs

It still seems to vary by market ( many factors including licensure in some
states), and by organization based on my observations. But I believe that
there is certainly a trend toward certification. At my organization they
require certification for any consideration, and also education to meet
CLIA.   But you are likely to get many different opinions on your question.




Joelle Weaver MAOM, HTL (ASCP) QIHC
 
 From: jon2038...@maricopa.edu
 Date: Thu, 22 Aug 2013 16:43:10 -0700
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] Uncertified Histotechs
 
 Is getting a job as an uncertified histotech a thing of the past? I have a
friend who has been a very skilled histotech for many years and they have
been looking for a job for about a year now. Is this due to bad interviewing
or a lack of certification? I'm curious to see if this has happened to other
people. They have applied at hospitals and bigger labs such as Caris. Im not
asking for a job lead for them I'm just more curious if certification has
become a prerequisite now.
 ___
 Histonet mailing list
 Histonet@lists.utsouthwestern.edu
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet
 
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


This message, and any included attachments, are from Virtua Health or its
related affiliates and is intended only for the addressee(s). The
information contained herein is privileged, proprietary or may include
confidential information and/or protected patient health information. Any
unauthorized review, forwarding, printing, copying, distributing, or
otherwise disseminating or taking any action based on such information is
strictly prohibited. If you have received this message in error, or have
reason to believe you are not authorized to receive it, please delete this
message promptly and notify the sender by e-mail with a copy to
issecur...@virtua.org. 

Thank you



___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

AW: [Histonet] EDTA decal

[Gudrun Lang]

We experienced different IHC staining qualities on bone marrow, that was
decalcified with EDTA or formic acid. I remember, our CD38 antibody was
always very weak after EDTA and made no problems after formic acid.
I think it is very difficult to make an over-all-statement for all
antigens/all antibodies. 
Gudrun 



-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Teri
Johnson
Gesendet: Freitag, 16. August 2013 19:54
An: 'cfors...@umn.edu'
Cc: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] EDTA decal

Hi Colleen,

I would say it's unusual, but not completely impossible that EDTA has
interfered with your IHC.  We had that problem with demonstrating
B-galactosidase in mouse bones. If we decalcified it in EDTA after whole
mount staining in X-gal, the blue staining was removed. But if we
decalcified it in formic acid, the stain was retained.

I think a few years ago, Biogenex had a room temperature antigen retrieval
solution for acid decalcified bone (not the same as your situation, I know).
But I think it was largely an alkaline solution you let the slides sit for
maybe 30 minutes prior to staining. Might be worthwhile to try a different
retrieval method for these and see what happens.

Otherwise, are you sure you are using a clone that reacts in mouse tissue?
We use Rabbit monoclonal SP6.

Teri Johnson
Manager, Histology
GNF - San Diego, CA
858-332-4752

___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet