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?


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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




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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

 

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[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

 

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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??

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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
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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
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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.
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[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





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[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

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[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

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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
Sent: Wednesday, 29 March 2023 4:51 AM
To:
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
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[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

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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


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[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

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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
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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
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[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

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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



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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

 

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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
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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
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[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



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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
www.caremountmedical.com



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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
>
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[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

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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

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[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

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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
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you are hereby notified that any dissemination, distribution or copying of
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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.
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[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

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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
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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
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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

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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
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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

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[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)

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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.
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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





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[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

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[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

 

 

 

 

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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.
>
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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 
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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

 

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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

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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


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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
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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

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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




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[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

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[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

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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







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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
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[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

 

 

 

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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
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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
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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

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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




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[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

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[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

 

 

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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
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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

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