Re: [Histonet] Plastic coverslip Loosening

2016-03-22 Thread Helen Fedor via Histonet
Thank you for your help. This is encouraging. 

Helen

-Original Message-
From: Glover, Kimberly [mailto:kimberly.glo...@polysciences.com] 
Sent: Tuesday, March 22, 2016 11:38 AM
To: Helen Fedor 
Cc: histonet 
Subject: RE: Plastic coverslip Loosening

Helen,

Submerge the slides with coverslip loosely attached in fresh xylene to 
reactivate the film resin. Allow to sit in xylene for approximately 1 minute. 
Afterwards remove slide and press down on a paper towel. This action will allow 
the film to reattach to the slide while pressing out any excess xylene from the 
slide. 

As for the film (containing tissue) that has completed detached from the slide, 
place the film against a clean slide and place in a slide basket. Submerge 
slide in fresh xylene and proceed with same procedure above. 

Sometimes the issue of loosening or lifting of film is seen over time when the 
coverslipped slides are placed in temperature/humidity conditions that are 
contrary to the manufacturer's recommendations. Check packaging for storage 
conditions recommended.

I hope this helps.

Kimberly A. Glover 
Life Sciences Product Manager
Polysciences, Inc.
400 Valley Road
Warrington, PA 18976
www.polysciences.com




-Original Message-
From: Helen Fedor [mailto:hfe...@jhmi.edu] 
Sent: Tuesday, March 22, 2016 11:06 AM
To: Glover, Kimberly
Subject: RE: Plastic coverslip Loosening

The film.

-Original Message-
From: Glover, Kimberly [mailto:kimberly.glo...@polysciences.com] 
Sent: Tuesday, March 22, 2016 10:25 AM
To: Helen Fedor 
Subject: RE: Plastic coverslip Loosening

Are you using plastic coverslipping film or the plastic coverslips?

Kimberly

-Original Message-
From: Helen Fedor via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Tuesday, March 22, 2016 9:45 AM
To: HISTONET (histonet@lists.utsouthwestern.edu)
Subject: [Histonet] Plastic coverslip Loosening

Hello, I did try and search the archives to see if there was any advice on this 
but didn't find an answer.

Do you know of a good way to fix H&E slides that had plastic coverslips where 
the coverslip is lifting from the slide and in some cases coming completely 
off? In all cases the tissue is on the coverslip.

Thanks in advance.



Helen L. Fedor


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[Histonet] Plastic coverslip Loosening

2016-03-22 Thread Helen Fedor via Histonet
Hello, I did try and search the archives to see if there was any advice on this 
but didn't find an answer.

Do you know of a good way to fix H&E slides that had plastic coverslips where 
the coverslip is lifting from the slide and in some cases coming completely 
off? In all cases the tissue is on the coverslip.

Thanks in advance.



Helen L. Fedor


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Re: [Histonet] MTA-1 Microarrayer

2015-05-04 Thread Helen Fedor
Hello Bernice, we have tried out several and found that "enercell" works. 
CR2450 their number is 23-808.

WE might have found them on amazon.

Cannot remember.
Helen L. Fedor 

Oncology Tissue Services, Manager
Johns Hopkins University
411 N. Caroline St 
Room 310 Basement| Bond St Annex Building
Baltimore, MD | 21231

410-614-1660

http://tmalab.jhmi.edu/






-Original Message-
From: Bernice Frederick [mailto:b-freder...@northwestern.edu] 
Sent: Monday, May 04, 2015 10:53 AM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] MTA-1 Microarrayer

Hello all,
Anybody have any idea where to get the batteries for this arrayer? Regular 2450 
batteries are too thick. Batteries plus was a no go. See some on Amazon but not 
sure if they are thin enough.
Thanks,
Bernice

Bernice Frederick HTL (ASCP)
Senior Research Tech
Pathology Core Facility
Robert. H. Lurie Cancer Center
Northwestern University
710 N Fairbanks Court
Olson 8-421
Chicago,IL 60611
312-503-3723
b-freder...@northwestern.edu

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RE: [Histonet] TMA Arrayers

2015-04-30 Thread Helen Fedor
Hello, We have the Pathology Devices Semi automated units. We are happy with 
them. Our decision to keep with the less automated units are largely due to our 
work flow. We make TMA's for customers. But the customer has to do all of the 
work in the design and data entry into out TMAJ Database. I think all of the 
automated units have data input features that are not compatible with this work 
flow.



Good luck in choosing.

Helen L. Fedor 

Oncology Tissue Services, Manager
Johns Hopkins University
411 N. Caroline St 
Room 310 Basement| Bond St Annex Building
Baltimore, MD | 21231

410-614-1660

http://tmalab.jhmi.edu/






-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sally Ann Drew
Sent: Thursday, April 30, 2015 1:36 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] TMA Arrayers

I was wondering how many people have semi-automated or automated tissue 
microarray equipment? I am especially interested in those who graduated from 
manual methods to the automated and how they reviewed and ranked the limited 
options out there.

Thank you!
_Sally

Sally Ann Drew, MT(ASCP)
UWSMPH-Dept. of Pathology 
TRIP Lab Manager 
Translational Science BioCore
CSC, L5/181
608.265.1093



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[Histonet] Vacuum processing of biopsies.

2015-03-31 Thread Helen Fedor
Hello all, I would be interested in pros and cons of using one of the Rapid 
vacuum processing units for processing of biopsy tissues.  Is this technology 
improving the processing?  Any specific problems that may be occurring.


Thanks in advance.

Helen L. Fedor

Oncology Tissue Services, Manager
Johns Hopkins University
411 N. Caroline St
Room 310 Basement| Bond St Annex Building
Baltimore, MD | 21231

410-614-1660

http://tmalab.jhmi.edu/




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RE: [Histonet] Cryosectioning undecalcified bone

2015-02-05 Thread Helen Fedor
Hello, I believe that the products for the CryoJane tape transfer are still 
available from Fisher.


Helen L. Fedor 

Oncology Tissue Services, Manager
Johns Hopkins University
411 N. Caroline St 
Room 310 Basement| Bond St Annex Building
Baltimore, MD | 21231

410-614-1660

http://tmalab.jhmi.edu/




-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth 
Chlipala
Sent: Thursday, February 05, 2015 1:08 PM
To: Orla M Gallagher; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Cryosectioning undecalcified bone

Orla

There is an article in the Journal of Histotechnology from a while ago from 
John Tarpley that addressed methods for this.  I have a pdf of it and I will 
send in another e-mail.

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 Orla M Gallagher
Sent: Thursday, February 05, 2015 10:56 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Cryosectioning undecalcified bone

Dear Histonetters,

We would like to develop cryosectioning for undecalcified mouse and rat bones. 
Previous attempts to use the CryoJane system from Instrumedics with an old 
Bright cryostat and solid tungsten carbide blade a few years ago didn't result 
in successful reproducible sections  - marrow without bone or bone without 
marrow on the slide, in spite of various freezing and prep.
protocols used by my colleague. I suspect the old cryostat and blade weren't 
helping either.

Have you any recommendations on the best cryostat to use to do this? We'd like 
to also use the cryostat for standard soft tissue sectioning. I've seen Leica 
mentioned in a few papers relating to CryoJane.

There appears also to be a tape transfer method from Kawamoto's Section-Lab Co. 
Ltd. (i...@section-lab.jp)

Does anyone use this method or know whether the consumables are still available 
for purchase, as the website seems dormant?

Thanks for any advice,
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.

http://www.shef.ac.uk/faculty/medicine-dentistry-health/2.9274/emailwording

http://www.sheffield.ac.uk/humanmetabolism/greenimpact


http://www.sheffield.ac.uk/visitors/mapsandtravel
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[Histonet] RE: Cassette Labeler

2015-01-27 Thread Helen Fedor
Hello, we recently purchased the Primera unit from Creative waste solutions.
http://www.cwsincorp.com/

we like it. I would be happy to discuss it further if you like.


Helen L. Fedor 

Oncology Tissue Services, Manager
Johns Hopkins University
411 N. Caroline St 
Room 310 Basement| Bond St Annex Building
Baltimore, MD | 21231

410-614-1660

http://tmalab.jhmi.edu/






-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Debbie Granato
Sent: Monday, January 26, 2015 12:07 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Cassette Labeler

Our lab is currently looking into purchasing a cassette and slide labeler.

We are a small lab and are looking for a stand- alone unit with the flexibility 
to be used with the computer or bar code scanner at a later date.

I would appreciate any feedback or suggestions for any model that may fulfill 
our needs.

Thank you,

Debbie Granato HT(ASCP)


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RE: [Histonet] Re: Histonet Digest, Vol 130, Issue 16

2014-09-18 Thread Helen Fedor
We had the PTL, which is a little more cumbersome. We are now using the BBP33, 
and are very happy with it.


Helen L. Fedor 

Prostate Tissue Bank, Manager
Oncology Tissue Services, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

410-614-1660 (Marburg)
443-287-7338 (Bond St.)

http://tmalab.jhmi.edu/
http://prostatebiorepository.org/





-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Dorothy Hu
Sent: Thursday, September 18, 2014 3:09 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Re: Histonet Digest, Vol 130, Issue 16

Hi Dear Histoneters,
Can you tell me which kind of Brady labeler you are using? I want to get one. 
Do you have catalog number? Many thanks.
Dorothy
MGH
Endocrine histocore

>
>
> I use the Brady system and I like it a lot.  I work in a small animal 
> diagnostic lab and will have 60-80 blocks on most days.  It saves me 
> time and even better I don't have to read my writing in the morning.  
> I use it for the cassettes and slides mainly but I label reagents I mix with 
> it to.
> I find it very versatile.
> Roberta Horner HT/HTL
> Animal Diagnostic Lab
> Penn State University
>
>
> Subject: [Histonet] Slide and cassette printers?
>
> What are all of you using these days?  What would be good for a small 
> histo lab that has the potential to grow?  (I am already aware of what 
> Leica and Thermo have from the archives here, but I am still 
> interested in your opinions, of course.)  Has anyone tried the 
> printers, labels and attachment machine from Brady?
>
> Thank you all so much,
> Kathleen
>
>
> Principal Lab Technician
> Histopathology Lab
> Office of Translational Sciences
> Rutgers, the State University of NJ
> 41 B Gordon Road
> Piscataway, NJ 08854
> (848) 445-1443
> FAX (732) 445-6905
>
>
>
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RE: [Histonet] Slide and cassette printers?

2014-09-17 Thread Helen Fedor
We are also using the Brady labeler. It is versatile, easy to use, they have 
great tech support. We are not using the cassette label attaching machine. For 
the cassette printer we are using the Primera. The Tech support there is also 
great.


Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Roberta Horner
Sent: Wednesday, September 17, 2014 8:20 AM
To: Kathleen Roberts; histonet
Subject: RE: [Histonet] Slide and cassette printers?

I use the Brady system and I like it a lot.  I work in a small animal 
diagnostic lab and will have 60-80 blocks on most days.  It saves me time and 
even better I don't have to read my writing in the morning.  I use it for the 
cassettes and slides mainly but I label reagents I mix with it to.  I find it 
very versatile.
Roberta Horner HT/HTL
Animal Diagnostic Lab
Penn State University

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kathleen Roberts
Sent: Tuesday, September 16, 2014 5:05 PM
To: histonet
Subject: [Histonet] Slide and cassette printers?

What are all of you using these days?  What would be good for a small histo lab 
that has the potential to grow?  (I am already aware of what Leica and Thermo 
have from the archives here, but I am still interested in your opinions, of 
course.)  Has anyone tried the printers, labels and attachment machine from 
Brady?

Thank you all so much,
Kathleen


Principal Lab Technician
Histopathology Lab
Office of Translational Sciences
Rutgers, the State University of NJ
41 B Gordon Road
Piscataway, NJ 08854
(848) 445-1443
FAX (732) 445-6905

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[Histonet] RE: rolling sections

2014-06-12 Thread Helen Fedor
Hi, I think that it is not necessary to actually get them to roll. We just 
collect all of the sections and put them into the tube. Scrunched, not rolled.

Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Roberta Horner
Sent: Thursday, June 12, 2014 10:42 AM
To: Histonet (histonet@lists.utsouthwestern.edu)
Subject: [Histonet] rolling sections

I have some researchers that want to do PCR.  They want 10 - 10u sections in a 
micro-centrifuge tube.  The only way to get the sections in the tube is for the 
sections to roll.  How do you get sections to roll when you want them to roll?  
I've tried room temperature, on ice, brand new sharp blade, dull blade and I 
can still get some really nice ribbons.  When I want a thick ribbon it will 
roll, darn that Murphy and his laws.
Roberta Horner
Animal Diagnostic Lab
Penn State University
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[Histonet] Temporary position

2014-01-08 Thread Helen Fedor
Hello, We are in search of a temporary histo-tech or histologist to work in our 
OTS Core lab. Primarily we do research work to support the efforts of the 
Cancer Center Investigators. A majority of the work is on animal tissues.
Gross, Process, Embed, section and do recuts, i.e., lining up blocks.  The lab 
also makes TMA's and coring paraffin blocks for RNA and DNA extractions. Please 
contact me if you are interested, or know someone that may be interested in 
this opportunity.

Thanks,

Helen


410.614.1660

http://tmalab.jhmi.edu/
http://prostatebiorepository.org/

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[Histonet] embedding cells in parrafin #2

2013-12-02 Thread Helen Fedor
Making cell tissue blocks

Cells: 
.   Harvest and fix in formalin* for at least 3hrs or overnight (10 fold 
more formalin)
.   HAVE A NICE VISIBLE SIZE PELLET (1 confluent T75 minimum or 1 T175)
.   Remove formalin* and wash 2x's in cold 1x PBS
.   Cover cells with some 1x PBS and refrigerate

Materials for blocks:
1.  Toothpick or 20ul pipette tip
2.  Agarose  (2% solution agarose with 1xPBS)
ultra pure agarose cat#15510-027 from core

3.  Cells in 1x PBS
4.  0.5ml microtubes (sterile)
5.  Tissue cassettes (1 or 6 compartiment cassettes -Electron Microscopy
 Science 800-523-5874 cat.# 70078-W)  
6.  New razor blades
7.  Clean cutting board
8.  Container to submerge cassette into with enough 1x PBS to cover 
cassettes

Method:
.   Suspend cells in 1xPBS - use just enough to pipette cells (1:1 dilution 
is MAX)
.   Make 2% agarose solution - keep in water bath (42C) so doesn't solidify
.   Make 200ul agarose plug in 0.5ml tube and wait 3 minutes to solidify 
.   Add *120ul of cell solution to .5ml tube and then add *150ul of agarose 
and pipette up/down ~3 times to mixed well - CUT END OF PIPETTE TIP OFF JUST A 
LITTLE SO YOU DON'T DESTROY CELL IF NECESSARY
.   Add toothpick and let sit 5 minutes
.   Remove plug with toothpick and cut into sections (if necessary) with 
new blade on clean surface and add to cassette
.   If you have extra cell plugs then store then in 1x PBS in frig.

* Varies according to how much cell suspension you have. ALWAYS add at least 
30ul more of agarose in order for it to make a nice plug.

IMPORTANT NOTES:
.   ALWAYS be sure to label your cassette with an experiment number because 
that's what the slides will be labeled with.
.   Be SURE to write on BOTH the top of the cassette and down the sides 
what cell type is in each compartment (if using a 6 compartment cassette)
.   Tell the histologist what you fixed your cells in - if fixed in 
formalin, ethanol, paraformaldehyde, etc. - this is vital for the 
processing/embedding process
.   ALWAYS put control cell types in the block  - +/-
.   ALWAYS prepare your block in an asymmetric pattern THIS IS A MUST!! or 
you will not be able to identify which cell type is placed where on the slide
Helen L. Fedor 

Prostate Tissue Bank, Manager
Oncology Tissue Services, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

410.614.1660

http://tmalab.jhmi.edu/
http://prostatebiorepository.org/



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mike Tighe
Sent: Monday, December 02, 2013 12:06 PM
To: histonet@lists.utsouthwestern.edu (histonet@lists.utsouthwestern.edu)
Subject: [Histonet] Embedding cells in Paraffin

Has anyone tried to embed cells grown in tissue culture? I am trying to put 
some tissue culture cells through same stress as tissue would go through. 
Fixation, dehydration, and heat. Any ideas? I could re-suspend in OCT and then 
fix for extended time with NBF but that doesn't quite seem fair.



Thanks for any ideas!
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RE: [Histonet] The Good Old Days...

2013-09-20 Thread Helen Fedor
You can tell it is Friday.

:)

Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cristi Rigazio
Sent: Friday, September 20, 2013 1:40 PM
To: Davis, Cassie
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] The Good Old Days...

Hear hear!  I agree and was just saying I love hearing the stories (although I 
am not young).  Thanks all for sharing these memories and lessons!

Sent from my iPhone

On Sep 20, 2013, at 10:13 AM, "Davis, Cassie"  wrote:

> I enjoy hearing sincere reminiscing...Even though us "kids" don't know how 
> good we have it, some of us enjoy having an "old tech" beside us on the 
> bench. I find weeding through the sarcasm can be profitable and in doing so 
> have learned so much. What the old techs did on a daily basis, we only did in 
> the practice lab and when the automated instruments and pre-made solutions 
> that we have come to rely on fail, experience is so very valuable. Only by 
> their blood, sweat and tears have we benefitted however, we have so far to 
> go, let's do it together.
> 
> Cassandra Davis
> cda...@che-east.org
> 302-575-8095
> 
> 
> 
> 
> 
> Confidentiality Notice:
> This e-mail, including any attachments is the property of Catholic 
> Health East 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.
> 
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RE: [Histonet] Re: Two Patient Identifiers on slides

2013-08-01 Thread Helen Fedor
Hello, I may not have read every e-mail on this discussion but would like to 
chime in here. I was just having this discussion with someone yesterday. At 
large research hospitals that have various goals in our mission statements, we 
are concerned about continuously improving the process to minimize any errors 
in patient care. But at the same time we also are using these same cases for 
research. Our IRB's allow us to use the Pathology Archives as a bank of tissues 
and slides. If the names of patients were on the slides and or blocks that 
would really make it impossible to do Specimen based research. We really 
couldn't use any of those materials for research. It has been a process here at 
Hopkins to try and keep both of these things moving forward, Improving patient 
care and maintaining access to specimens for research.


Helen L. Fedor 

Prostate Tissue Bank, Manager
Oncology Tissue Services, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

410.614.1660

http://tmalab.jhmi.edu/
http://prostatebiorepository.org/





-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bob Richmond
Sent: Wednesday, July 31, 2013 7:07 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Re: Two Patient Identifiers on slides

Realizing we're talking about regulator compliance rather than good sense here 
- nonetheless let a pathologist observe that having the patient's name on the 
slide is a very important defense against mixing up cases at sign-out. Small 
labs still usually have no identifier on the slide other than a hand-scribbled 
accession number (two of the three small labs I'm working in at the moment 
don't - both CAP accredited), and it's easy for the pathologist to misread the 
label. I saw a mixup like this just a few days ago - it wasn't my case, but I 
should have caught the misreading when I reviewed the case, which involved a 
squamous carcinoma that wasn't.

Good management, bad medicine.

Bob Richmond
Samurai Pathologist
Maryville TN
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RE: [Histonet] Block labeling

2013-06-24 Thread Helen Fedor
Hello, we are using the labels from the "Brady" label maker with good success. 
They are extra sticky and you can find some that will stick to paraffin. We use 
ones that qualify for freezing since they need to be safe on the ice tray.

http://www.bradycorp.com/

Helen


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Patrick Laurie
Sent: Monday, June 24, 2013 12:00 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Block labeling

Hi Everyone,

What is the best way to label blocks that are already embedded?  We have a 
large control bank that we are trying to relabel and make search able, however, 
most marking pens don't write well on paraffin covered blocks, even when we 
scrape them off.  Pencils work, not always the most legibly, so I was wondering 
if there is a clever solution that someone might have found.  We've tried 
putting stickers or labels on the back, but that falls right off.

Thanks in advance for any suggestions.

-- 

Patrick Laurie(HT)ASCP QIHC

Histology Manager

Celligent Diagnostics, LLC

101 East W.T. Harris Blvd  | Suite 1212 | Charlotte, NC 28262

Work: 704-970-3300  Cell: 704-266-0869
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[Histonet] RE: Control Storage

2013-06-12 Thread Helen Fedor
Hello, We are storing our unstained slides at -20 in Ziploc bags. Our TMA 
blocks are being stored at 4 degrees.

Helen L. Fedor 

Oncology Tissue Services, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

410.614.1660

http://tmalab.jhmi.edu/
http://prostatebiorepository.org/



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth 
Chlipala
Sent: Wednesday, June 12, 2013 11:56 AM
To: Herring, Colleen; Histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Control Storage

We store our blocks at room temp, our IHC slides are stored in the fridge.

Liz

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

Ship to address:

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

From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Herring, Colleen 
[colleen_herr...@bshsi.org]
Sent: Wednesday, June 12, 2013 9:48 AM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Control Storage

Does anyone have any ideas or suggestions about the storage of blocks and 
precut slides for immunos?


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[Histonet] RE: Out of my comfort zone...

2013-04-03 Thread Helen Fedor
We have been using store bought gallons of distilled water in our water baths. 
This water has been boiled so enzyme activity should be absent.


Helen

410.614.1660

http://tmalab.jhmi.edu/
http://prostatebiorepository.org/



Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sue Hunter
Sent: Wednesday, April 03, 2013 10:55 AM
To: Sarah Dysart; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Out of my comfort zone...

Hello
Just wanted to add one more thing - we actually use a dedicated pyrex dish 
(maybe 6x10 inches) for our water bath for RNA sections.  We use warm tap 
water, but you can put it in the microwave for a short bit if it needs to be 
warmer.  You can spray the dish with RNAse away and wipe before filling with 
water.
Sue

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sarah Dysart
Sent: Tuesday, April 02, 2013 5:36 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Out of my comfort zone...

So...I have been asked to do some micro-dissection on some slides and then do 
downstream RT/PCR on them.  My molecular knowledge doesn't go much out of the 
world of IHC so...here is my question...

Has anyone ever substituted Citrate Buffer pH6 (or whatever HIER solution you 
are using) for proteinase K for use in RNA isolation and then later PCR?  Does 
this work?  The main question is will the HIER step take off the formalin 
linkage from the nucleic acids, or just the protein?

One last thing is what else goes into these solutions other than Citrate Buffer 
and Tween?  I haven't made it up in forever, I have just been ordering it from 
companies...I know...lazy...

Thanks

Sarah Goebel-Dysart, BA, HT(ASCP), QIHC (ASCP) Histotechnologist Mirna 
Therapeutics
2150 Woodward Street
Suite 100
Austin, Texas  78744
(512)901-0900 ext. 6912

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[Histonet] RE: minus 80 freezer safe time in power outage

2013-03-29 Thread Helen Fedor
Hello,  I think it would depend on what you think the malfunction is due to. If 
you do not have a solution in sight. I would start thinking about moving them 
immediately. RNA is sensitive to this type of warming.  How long is the 
transfer going to take? If they already up to -40 they are going to be to -30 
when they get to their new destination. And that is a problem.

Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Morken, Timothy
Sent: Friday, March 29, 2013 11:16 AM
To: Histonet
Subject: [Histonet] minus 80 freezer safe time in power outage

Hi all, For a minus 80 freezer what would you consider a "safe" time period 
that it can go without power before the temperature gets too high and contents 
need to be transferred (assuming door stays closed)?  I'm thinking -40 would be 
a cutoff temp to move samples. Anyone else think differently about that?

Thanks for any info!


Tim Morken
Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of 
Pathology UC San Francisco Medical Center
505 Parnassus Ave, Box 1656
Room S570
San Francisco, CA 94143

(415) 353-1266 (ph)
(415) 514-3403 (fax)
tim.mor...@ucsfmedctr.org

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[Histonet] RE: Tissue Microarray

2013-01-23 Thread Helen Fedor
Hello Donna, We have two different instruments, and they each have value. But 
if you are only interested in making small arrays for control blocks  then you 
wouldn't need to spend the amount of money that the larger Arrays would cost. 
http://www.pathologydevices.com/TMArrayer.htm
this company sells both kinds and also sells the needles.
The former company Beecher is now in Estonia under http://www.estigen.com/


I would be happy to speak to you about other small models if you are interested 
or would like to discuss these instruments.


Helen L. Fedor 

Prostate Tissue Bank, Manager
Oncology Tissue Services, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

410.614.1660

http://tmalab.jhmi.edu/
http://prostatebiorepository.org/


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Willis, Donna G.
Sent: Wednesday, January 23, 2013 1:20 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Tissue Microarray

I need to purchase a Tissue Microarray instrument.  Would folks please give me 
their preferences and why.

Thanks,

Donna Willis, HT/HTL (ASCP)
Anatomic Pathology Manager
Baylor University Medical Center-Dallas
ph. 214-820-2465 office
ph. 214-725-6184 mobile
donna.wil...@baylorhealth.edu

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RE: [Histonet] Validation of a new tissue processor

2012-11-28 Thread Helen Fedor
You will also need to validate all of the IHC stains and special stains that 
you do on these tissues. You can take samples from each tissue type and run 
them in parallel on both machines. Then test all of your special stains and 
antibodies that you have in your inventory.

:)
Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Wednesday, November 28, 2012 4:47 PM
To: Howe, Helena; 'histonet@lists.utsouthwestern.edu'
Subject: Re: [Histonet] Validation of a new tissue processor

Using at least 25 different types of tissues, prepare parallel almost identical 
pairs of slices for each tissue and process them simultaneously in your new 
tissue processor and in the old one.
Prepare H&E sections and give them to the pathologists concealing the used 
tissue processor. The pathologists should decide if there are differences 
between the pairs of sections.
The pathologist's opinion should be registered in the QC file and that will be 
your validation.
René J.

From: "Howe, Helena" 
To: "'histonet@lists.utsouthwestern.edu'"  
Sent: Wednesday, November 28, 2012 2:57 PM
Subject: [Histonet] Validation of a new tissue processor

Does anyone know what the CAP is requiring to validate a new tissue processor?
Thank You
Helena Howe
ProMedica Laboratories
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RE: [Histonet] Need better results from older archival slides?

2012-11-19 Thread Helen Fedor
Agreed!


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Monday, November 19, 2012 11:16 AM
To: Rob Day; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Need better results from older archival slides?

Rob:
If you are really seeking for a confirming study, publish your data and 
probably there will be a confirmation study.
 
I went to the website you indicated and it seems to me that your original 
posting is just a "subtle and delicate" advertisement.
 
Just my opinion otherwise, why don't you tell us all what is that you do to 
obtain your rejuvenating results? 
René J.



From: Rob Day 
To: histonet@lists.utsouthwestern.edu 
Sent: Monday, November 19, 2012 11:03 AM
Subject: [Histonet] Need better results from older archival slides?


Hello,

Having problems getting good Immuno staining and in-situ hybridization results 
with older slides?

I am writing to see if histonet can suggest any scientists who may be 
interested in helping us with a study related to a  "slide rejuvenation" 
process we are developing.

We think  we have found a way to significantly improve the antigenicity of 
proteins in older archival FFPE preserved slides. Initial tests seem to 
indicate that our process can allow slides of 5-10 years age, possibly older, 
to respond to IHC analysis as if they were cut that day. The process may have 
additional uses and benefits too, including better preservation of cut 
sections, enhanced ISH results, stabilization of labile proteins and 
standardization / optimization of IHC staining protocols. We are excited about 
this development, but we are seeking some independent labs to help us verify 
our findings.

Please feel free to call me to discuss, or pass on this email to any 
investigators you may know of who are exploring ways to make better use of 
older archival slides.

Folio Biosciences is a provider of biosamples and a commercial research lab 
based in Columbus Ohio. See: http://www.foliobio.com/.

Regards,

Rob Day.






Rob Day
Business Development 
Folio Biosciences
1476 Manning Pkwy, Powell, Ohio 43065
Direct Line: (614) 407-4547 | Main Office Phone: (614) 846-2809 |  Fax: (877) 
591-1815
http://foliobio.com

www.linkedin.com/in/robdaybiotech

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[Histonet] Bliss Imaging system

2012-07-26 Thread Helen Fedor
Hello, We have a Bliss imaging system that we are no longer using. I has a 
great Zeiss axiophot microscope with high quality lenses. I believe that 
Olympus is now the company that has taken over the Bacus Labs Imaging system. 
Please contact me if you have a desire to acquire this system from us or know 
of anyone who might be interested in it..

Thanks for your consideration,

Helen L. Fedor

Prostate Tissue Bank, Manager
TMA Core Lab, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

410.614.1660

http://tmalab.jhmi.edu/
http://prostatebiorepository.org/

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[Histonet] RE: Beecher

2012-07-11 Thread Helen Fedor
http://www.pathologydevices.com/TMArrayer.htm

You can contact Ron Gebing at Pathology Devices.


Helen
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bernice 
Frederick
Sent: Wednesday, July 11, 2012 12:14 PM
To: Fellow HistoNetters
Subject: [Histonet] Beecher

All
,I know this came up before,but who took over Beecher? We need to order some 
more punchers.
Thanks,
Bernice

Bernice Frederick HTL (ASCP)
Senior Research Tech
Pathology Core Facility
ECOGPCO-RL
Robert. H. Lurie Cancer Center
Northwestern University
710 N Fairbanks Court
Olson 8-421
Chicago,IL 60611
312-503-3723
b-freder...@northwestern.edu

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RE: [Histonet] Billing 88342

2012-07-03 Thread Helen Fedor
That is so true.!

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Paula Pierce
Sent: Tuesday, July 03, 2012 1:36 PM
To: Jay Lundgren; Victor A. Tobias
Cc: HISTONET
Subject: Re: [Histonet] Billing 88342

Where is the like button?


Thumbs up!

 
Paula K. Pierce, HTL(ASCP)HT
President
Excalibur Pathology, Inc.
8901 S. Santa Fe, Suite G
Oklahoma City, OK 73139
405-759-3953 Lab
405-759-7513 Fax
www.excaliburpathology.com



 From: Jay Lundgren 
To: Victor A. Tobias 
Cc: HISTONET 
Sent: Tuesday, July 3, 2012 12:05 PM
Subject: Re: [Histonet] Billing 88342
 
     I am neither a lawyer nor a health care administrator, but, in my 
experience, the Pathologist picks the (hopefully) most diagnostic blocks from 
the multiblock cases and submits them for IHC.  If you do the requested IHC on, 
say, 4 blocks out of 30, you charge x4 for the technical fee.  After all, you 
are using 4 times the supplies (buffer, antibody, etc.).
     Before you hit the cash paying patient with a bill, their primary care 
provider should warn them what it's going to cost.
     I have seen a good Pathologist only select one block for IHC when the 
clinician previously informed him that the patient had no insurance and was 
paying out of pocket.
     I think it's interesting that people control *their own* health care costs 
when no insurance company or the government is involved.

                                           Sincerely,

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

On Tue, Jul 3, 2012 at 11:43 AM, Victor A. Tobias  wrote:

> Looking for other opinions from those who do consult/referral work.
>
> If a client sends in a request for a single antibody done on multiple 
> blocks on a single specimen, do you bill the client for each tech 
> component ? The client will do the interpretation.
>
> What happens in the above scenario if the request is to bill the patient?
> Knowing you get reimbursed for one, do you eat the other charges are 
> make the client select the one block?
>
> We have run numbers on potential lost revenue and the number is 
> significant.
>
> Victor
>
>
> Victor Tobias HT(ASCP)
> Clinical Applications Analyst
> Harborview Medical Center
> Dept of Pathology Room NJB244
> Seattle, WA 98104
> vtob...@u.washington.edu
> 206-744-2735
> 206-744-8240 Fax
> =
> Privileged, confidential or patient identifiable information may be 
> contained in this message. This information is meant only for the use 
> of the intended recipients. If you are not the intended recipient, or 
> if the message has been addressed to you in error, do not read, 
> disclose, reproduce, distribute, disseminate or otherwise use this 
> transmission. Instead, please notify the sender by reply e-mail, and 
> then destroy all copies of the message and any attachments.
>
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RE: [Histonet] TMA blocks

2012-02-20 Thread Helen Fedor
Hello Colleen, We use standard size molds (2.5cm X 3.5cm) that are a litte 
deeper (7mm) in our core to make the recipient blocks. 2.5cm X 3.5cm. and our 
TMA with the 1.0mm punch have 13 rows and 16 columns = 208 cores.  You can 
contact me if you want more information.


Helen L. Fedor 

Tissue Microarray Lab, Manager
Prostate Biorepository, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

410.614.1660

http://tmalab.jhmi.edu/





-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Colleen Forster
Sent: Monday, February 20, 2012 5:02 PM
To: Histonet
Subject: [Histonet] TMA blocks

Hey Histonetters!

We have the first generation of the Beecher machine, manual version. We would 
like to do a super TMA that would hold 180+1.0mm punches. When you do a single 
TMA the tissue area is 1x1 and out largest mold is 3x3. Has anyone ever trimmed 
the TMA's down and embedded 2 or 3 of them into one big block?

Thanks in advance!!

Colleen Forster
Bionet Histology Laboratory
U of MN, AHC

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RE: [Histonet] Processor Recomendations?

2012-01-31 Thread Helen Fedor
Where is the LIKE button?

Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Tuesday, January 31, 2012 3:31 PM
To: histonet@lists.utsouthwestern.edu; Elizabeth Cameron
Subject: Re: [Histonet] Processor Recomendations?

I used to work with 3 VIPs, one of which was 15 years old. I really doubt that 
any of your VIPs "could go at any time".
I suggest that the money you have invest in refurbishing your VIPs, a capital 
maintenance that the people from Sakura can do.
I think you should go this way before investing in another instrument.
René J.

--- On Tue, 1/31/12, Elizabeth Cameron  wrote:


From: Elizabeth Cameron 
Subject: [Histonet] Processor Recomendations?
To: "histonet@lists.utsouthwestern.edu" 
Date: Tuesday, January 31, 2012, 2:57 PM


Hi,
I am looking for a reasonably-priced tissue processor for a research facility.  
We only process mouse tissue (100-200/day), and rapid processing is not 
something that is necessary.  If possible, we would like the option of using 
isopropyl alcohol for processing.  We currently have 2 Sakura VIPs that we are 
happy with, however, they are old and could go at any time.  Any 
suggestions/recommendations would be appreciated.
Thank you!

Elizabeth M. Cameron, HT, QIHC (ASCP)
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[Histonet] RE: Casual Temp Position

2012-01-09 Thread Helen Fedor
Hello, We have a position available for a casual temp employee. This is a 
reference lab located in Baltimore MD, that primarily focuses on animal 
specimens. We will consider someone who has 2 years or more cutting experience. 
Please forward your resume to me.

thanks

Helen
410.614.1660

http://tmalab.jhmi.edu/ 
Helen



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Helen Fedor
Sent: Monday, January 09, 2012 1:37 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Casual Temp Position

Hello, We have a position available for a casual temp employee. This is a 
reference lab that primarily focuses on animal specimens. We will consider 
someone who has 2 years or more cutting experience. Please forward your resume 
to me.

thanks

Helen
410.614.1660

http://tmalab.jhmi.edu/

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[Histonet] Casual Temp Position

2012-01-09 Thread Helen Fedor
Hello, We have a position available for a casual temp employee. This is a 
reference lab that primarily focuses on animal specimens. We will consider 
someone who has 2 years or more cutting experience. Please forward your resume 
to me.

thanks

Helen
410.614.1660

http://tmalab.jhmi.edu/

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RE: [Histonet] to cut or not to cut mouse brains

2011-11-17 Thread Helen Fedor
When the blocks are not croyprotected you can cut them at warmer temperatures. 
But when they are cryoprotected with the 30% sucrose, they are soft at -18, 
that is why you need the colder temperature. they will compress at the warmer 
temperatures and give you a lot of wrinkles.

Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Grantham, 
Andrea L - (algranth)
Sent: Thursday, November 17, 2011 2:57 PM
To: McLaughlin, Terry
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] to cut or not to cut mouse brains

I'm interested in this answer because this has happened to me - but not always.
I usually cut brains at warmer temps. like -17 or -18 as opposed to -25 and 
they are usually handled in the same manner before being brought to the lab 
frozen in OCT.

Andi



On Nov 17, 2011, at 12:24 PM, McLaughlin, Terry wrote:

> Hello All,
> I am having trouble cutting frozen mouse brains and was wondering if 
> someone can offer some help. The mouse was perfused in 4% PFA , the 
> brain removed and kept in PFA for 22 hrs. We placed it in 30% sucrose 
> for 20 hrs, until the brain sank in this solution. It was frozen back 
> by using a culture plate floating in liquid nitrogen with OCT .
> The problem I am having is many folds, and air bubbles under the tissue.
> What I have done so far:
> Cut at temp of -25, then tried - 22 and -17.
> Cut at 10um, then tried 16-25um.
> Tried making the knife colder by adding dry ice to it for a few seconds.
> Added dry ice to sample for a couple seconds.
> The sample cuts fine, it appears to happen when picking up on a slide. 
> I tried picking up by starting at one side or end and letting tissue 
> spread onto slide. Did not work.
> Also tried gently laying slide on top of sample and removing 
> gently-still folds and air bubbles. Any help would be greatly 
> appreciated as we have 6 more to do.
> 
> Signed,
> Help!
> Terry Kokas
> 
> 
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> 


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[Histonet] RE: to cut or not to cut mouse brains

2011-11-17 Thread Helen Fedor
Hello Terry,

These are very difficult. The colder the better. (-27). keep the slides in the 
cryostat. Cut the section. Place section on a cold slide, and keep the slide in 
the croystat during the following procedure.

Hold slide in left hand and brush in right. Place finger under one edge of the 
section and it will start to thaw, anchoring it to the slide. Gently brush out 
folds as you continue to advance the thaw line with your fingers under the 
slide. It takes a bit of practice, and not really perfect. But it works.

Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of McLaughlin, 
Terry 
Sent: Thursday, November 17, 2011 2:25 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] to cut or not to cut mouse brains

Hello All,
I am having trouble cutting frozen mouse brains and was wondering if
someone can offer some help. The mouse was perfused in 4% PFA , the
brain removed and kept in PFA for 22 hrs. We placed it in 30% sucrose
for 20 hrs, until the brain sank in this solution. It was frozen back by
using a culture plate floating in liquid nitrogen with OCT . 
The problem I am having is many folds, and air bubbles under the tissue.
What I have done so far:
Cut at temp of -25, then tried - 22 and -17.
Cut at 10um, then tried 16-25um.
Tried making the knife colder by adding dry ice to it for a few seconds.
Added dry ice to sample for a couple seconds.
The sample cuts fine, it appears to happen when picking up on a slide. I
tried picking up by starting at one side or end and letting tissue
spread onto slide. Did not work.
Also tried gently laying slide on top of sample and removing
gently-still folds and air bubbles. Any help would be greatly
appreciated as we have 6 more to do.
 
Signed,
Help!
Terry Kokas
 
 
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[Histonet] RE: need help with TMA

2011-10-31 Thread Helen Fedor
Hello, Do 30 minutes at 37 degrees on a glass slide. take from oven and apply 
gentle pressure to center of block and slide complex, cool to RT and repeat 1 
or two times.

Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Patricia F Lott
Sent: Friday, October 28, 2011 1:13 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] need help with TMA

We have recently started doing tissue microarrays.  The problem we have is that 
some of the small cores drop out after the first few sections.

I think my problem is in the step where we are supposed to melt the donor and 
recipient blocks together.  We tried 5 minutes at 37 degrees, and then  minutes 
at 60 degrees x3, with cooling to room temp in between.

Any suggestions?

Thanks,
Patty Lott, UAB
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RE: [Histonet] RE: DAPI on thicker sections, problem with gradient

2011-10-06 Thread Helen Fedor
Hello, Maybe this method was meant to be used on Floating sections instead of 
mounted sections. That way the binding and penetration would go from both sides 
of the tissue.

Helen L. Fedor 

http://tmalab.jhmi.edu/



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of gayle callis
Sent: Thursday, October 06, 2011 12:00 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: DAPI on thicker sections, problem with gradient

Carmen, 

 

You wrote: 

 

Good morning:

 

I am Carmen Garcia from the Faculty of Medicine of the University of 

Valencia, Spain. I usually use your vectashield mounting medium for 

fluorescence with DAPI (H-1200) and I never have any problem.

But now I am working with 50um frozen sections and the problem that I 

have is that the DAPI staining does not arrive to the bottom of the 

section. Do you have any sugesstion of what can I do? (I already have 

the sections permeabilized with triton x-100) I have thougth to 

incubate with the mounting medium some hours at 4ºC or put some 

mounting medium in the antibody diluent and incubate...

 

any suggestion would be really welcome.

Thank you so much,

Kind regards,

 

Carmen

 


***

The problem you are experiencing is what we refer to as a DAPI staining
gradient.  Uneven staining occurs with the DAPI is not reaching all the
cells due to viscosity of the mounting media and thickness of the tissue
section.  

 

You did not say whether your thick sections have been fixed before
sectioning and staining for a tissue component, and then the DAPI - please
explain.  I suggest you do NOT use a mounting media with DAPI, but rather
make a DAPI solution which you can then apply to the section and incubate
mounting the coverslip with Vectashield that does not contain DAPI.  Using a
DAPI solution may shorten the time you now use, e.g. "some hours".You
can buy the DAPI, ready to use from Biogenex or other vendors, but making it
in house is simple. I would assume you could add Triton X to the DAPI
solution  to improve permeability and match what you have already done with
that detergent.  After DAPI staining, mount the coverslip using  VectaShield
without DAPI.  DAPI recipes can be found on the internet, just Google DAPI.
I found it on a Johns Hopkins webpage, but IHC world may have it too.
There are other sources of ready to use DAPI if you look.  

 

We have used Prolong Gold antifade reagent with DAPI,  and get a staining
gradient on 5 um sections.   This is very annoying, and probably caused by
the actual technique of cover slipping itself. We are now going to use
Biogenex ready to use DAPI before the Prolong Gold antifade reagent without
DAPUI.   Staining for a thin section takes 5 mintues at RT, so you should
try several 50 um  sections (non experimental) to determine your optimal
staining time.   It may take only 15 minutes although cold, 4C temperature
may slow down the penetration of the stain through the thicker section.
This way you can achieve even staining of your nuclei without long
incubations in the more viscious mounting media containing DAPI. 

 

Good luck, and let us know if you have success...

 

Gayle M. Callis

HTL/HT/MT(ASCP)

 

 

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[Histonet] RE: Paraffin sections for molecular assays

2011-10-04 Thread Helen Fedor
Hello, For DNA, we use a new blande and we wipe down microtome, handles, front 
of blade holder with 95% ethanol on a gauze sponge. For RNA,  use a new blade 
and wipe down all with RNAzap including the water bath and rinse with DEPC 
water. Use  clean water in bath, distilled will work or DEPC water. Depending 
on the stringency of the assay. You can sometimes get away with a little less. 
But some of the assays that are being run can cost up to $1,000.00. So the time 
spent cleaning up is justified. Make sure you are wearing gloves for both RNA 
and DNA, during the whole process, even when handling the slides .(including 
the process of labeling slides). I usually lay the slides out on pape rtowels 
on the lab bench while labeling them.

Helen Fedor
Johns Hopkins University
TMA and Prostate Spore Lab Manager
JHU SOM Uro/Pathology
600 N. Wolfe Street, Marburg Room 406
Baltimore MD, 21287-7065
410-614-1660 / Fax 410-614-3695
Pager 410-283-3419



From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] on behalf of Crowell, Thomas 
[thomas.crow...@novartis.com]
Sent: Tuesday, October 04, 2011 12:44 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Paraffin sections for molecular assays

Dear Histonetters,

Can you tell me what procedures you use to prevent DNA/RNA contamination of 
tissue sections when handling multiple samples.  Do you just wipe down blade 
holder and blade (or use a new blade) between samples, or do you use more 
stringent cleaning?


Thomas Crowell
Diagnostic Development
Novartis Molecular Diagnostics
45 Sidney Street
Cambridge, MA 02139
USA

Phone+1  617 8717460
Fax +1  N.A.
thomas.crow...@novartis.com<mailto:thomas.crow...@novartis.com>
www.novartis.com<http://www.novartis.com/>


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RE: [Histonet] Control Slides

2011-03-15 Thread Helen Fedor
Hello,  We have been storing our slides in small zip loc bags at -20 and have 
data suggesting that slides stored in this fashion are reasonably well 
preserved after 3- 5 years. This was done with TMA slides. Was not done for 
special stain slides but for IHC. 


Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of 
pru...@ihctech.net
Sent: Tuesday, March 15, 2011 8:56 AM
To: Harrison,Sandra C.
Cc: histonet@lists.utsouthwestern.edu; Amador, Amanda
Subject: RE: [Histonet] Control Slides


   I do not melt dry my cut control slides until i need to use them,=d
   i  do store them in boxes at 4c, i have recently run into this problem
   =th  blocks,  so  now i seal all my blocks with paraffin and store
   them in th�ridge as well.



   Patsy



    Original Message 
   Subject: RE:=istonet] Control Slides
   From: "Harrison, Sandra C." <[1]sandra.harris...@va.gov>
   Date: Mon,=rch 14, 2011 1:40 pm
   To: "Lee & Peggy Wenk" <[2]lpw...@sbcglobal.net>, "Amador, Amanda"
   &l=[3]aama...@ameripath.com>,&=;[4]histo...@lists.utso   uthwestern.edu>
   We  dry  the  control  slide at the same time tha=e dry the tissue
   being
   stained,  since  controls  are  supposed  to  be han=ed in the same
   manner as
   the test tissue. That being said, I guess if = were truly following
   that rule, we would cut the control on the same �y we cut the test
   sample. :-)
   "Other people say that, after   they  dry  the  slide,  they  dip  the slide 
in paraffin, to cover the
   tissu=
   so
   that  the  air  doesn't touch the tissue during the months before=e
   slide
   is
   used in a stain."
   -Original Message-
   From: [5]histone=boun...@lists.utsouthwestern.edu
   [[6]mailto:histonet-bounces@lists.utsouthwestern.e=]  On Behalf Of
   Lee &
   Peggy Wenk
   Sent: Thursday, March 10, 21 6:39 PM
   To: Amador, Amanda; [7]histonet@lists.utsouthwestern.edu
   Subject: Re: [Histone= Control Slides
   I've  noticed  that  our  spirochete  control  slides do=t stain as
   intense
   starting about 3 months after they are sectioned, =d that they stop
   staining by about 6 months. (This is with a silver s=in.)
   I've  talked  with  other  histotechs, and they say they've seen =e
   same
   phenomenon with AFB and Gram controls. (We use ours up too qui=ly -
   we
   never  have  6-12  months  old  control slides for these, so I can=
   attest
   to
   this. )
   Bancroft's  book  talks  about  this  phenome=n  with amyloid, and
   suggests
   that
   oxidation of the tissue (protein= due to exposure to air may be the
   cause.
   I'm  guessing  the  it  prob�ly applies to the precut microorganism
   control
   slides, too.
   We  only  cut  enough slides for 6 months, and date them. Other people
   say
   they
   put their cut control slides in a slide box with a lid after dryi=,
   and
   then place the box in the refrig, as cold slows down the che=cal
   change,
   and  the  lid  keeps  the moving air off the slide. Other =ople say
   that,
   after
   they  dry  the  slide,  they dip the slide in par�fin, to cover the
   tissue,
   so
   that  the  air  doesn't touch the tissue=ring the months before the
   slide
   is
   used in a stain. Just 3 sugge=ions to stop this problem.
   Peggy A. Wenk, HTL(ASCP)SLS
   Beaumont=spital
   Royal Oak, MI 48073
   ---=-
   From: "Amador, Amanda" <[8]aama...@ameripath.com>
   Sent: Wednesday, March 09, 2010:22 PM
   To: <[9]=sto...@lists.utsouthwestern.edu>
   Subject: [Histonet] Control Sl=es
   >  Is there guidelines for how long special stains controls a= good
   for
   once
   >  they  are  cut?  We have spirochetes for our Stei=r that is from
   2007
   and
   > we are having issues.
   >
   >=anda Amador, HT(ASCP)CM
   >  AmeriPath  |  Histology  Group Lead/Trainer=560 N Shadeland Ave,
   Suite
   A |
   > Indianapolis, IN 46219 | phon�17.275.8052 |
   > [10]aamador@=eripath.com; |
   > [12]www.AmeriPath.com;
   >
   > ___   > Histonet mailing list
   > [13]Histonet@lists.utsouthwestern.edu
   > [14]http://lists.utso=hwestern.edu/mailman/listinfo/histonet
   __=___
   Histonet mailing list
   [15]Histonet@lists.utsouthwestern.edu[16]htt   
p://lists.utsouthwestern.edu/mailman/listinfo/histonet
   _=
   Histonet mailing list
   [17]Histonet@lists.utsouthwestern=du
   [18]http://lists.utsouthwestern.edu/mailman/listinfo/histonet
   

References

   1. file://localhost/tmp/3D"ma   2. file://localhost/tmp/3D"mailto   3. 
3D"mailto:aama...@ameripath.com";
   4. 3D"mailto:histonet@lists.utsouthwestern.edu";
   5. 3D"mailto:histonet-boun...@lists.utsouthwestern.edu";
   6. 3D"mailto:histonet-bounc   7. 3D"mailto:histonet@lists.utsouth  

RE: [Histonet] RE: Beecher Instruments

2011-03-04 Thread Helen Fedor
We too have tried the Quick Ray, and I think it is fine if you are not in the 
business. But if you only make a few arrays at a time then it is really 
wonderful I think for making small control block TMA's it is perfect. You can 
cut down the size of the recipient blocks to be able to make several out of 
each of these. The resultant blocks cut like butter. You don't lose a single 
section and you can cut them easily at 3 microns.


Helen L. Fedor

Tissue Microarray Lab, Manager
Prostate Spore Lab, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

410.614.1660







From: Mark Tarango [mailto:marktara...@gmail.com]
Sent: Friday, March 04, 2011 4:20 PM
To: Helen Fedor
Cc: Barone, Carol; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] RE: Beecher Instruments

There is also the Sakuro Quick-Ray System.  I don't know if you've tried that 
already, but I've used it in the past and liked it.  
http://www.sakura-americas.com/products/tisstek-quickray.html The list price is 
$3950 for the whole kit and the recipient blocks are $68 each regardless of 
size (1 mm, 2 mm, 3 mm, or 5 mm).

Mark
On Fri, Mar 4, 2011 at 1:09 PM, Helen Fedor 
mailto:hfe...@jhmi.edu>> wrote:
Hello, There is a company that I just ran a few months ago. 
http://www.pathologydevices.com/TMArrayer.htm  they have a manual arrayer that 
is running about 30,000.00. A little more than the Beecher Arrayer but it does 
offer some improvements. They are also able to sell the Needles that will fit 
into the Beecher instrument. I have spoken to Ron Gebing at Pathology Devices 
and he is responsive and reachable.

I am very optimistic, He has many units in use in the U.S. and the world.



Helen L. Fedor

Tissue Microarray Lab, Manager
Prostate Spore Lab, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

410.614.1660





-Original Message-
From: 
histonet-boun...@lists.utsouthwestern.edu<mailto:histonet-boun...@lists.utsouthwestern.edu>
 
[mailto:histonet-boun...@lists.utsouthwestern.edu<mailto:histonet-boun...@lists.utsouthwestern.edu>]
 On Behalf Of Barone, Carol
Sent: Friday, March 04, 2011 3:56 PM
To: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>
Subject: [Histonet] Beecher Instruments
Importance: High

Histonetters...It has been a longtime concern of mine that the only
company offering TMA instrumentation was Beecher Instruments.and the
wait list for those, was totally unacceptable. It came to my attention
today, from a reliable source at a university that Beecher has gone
out of business...( after buying Chemicon's arrayer...and taking away
the only real alternative). I have heard from  the same reliable source
they may be purchased by a foreign investor...my prayer is that perhaps
that could be Leica! Does anyone have more info on this? I wonder what
this means for those who already have one.at least I am on the wait
list!  We have tried several manual systems from molds to punchesand
there seems no real competition from these, which are difficult to
control or expensive to use.  What is the word out theresuggestions
for us "wait-ors". CB
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[Histonet] RE: Beecher Instruments

2011-03-04 Thread Helen Fedor
Hello, There is a company that I just ran a few months ago. 
http://www.pathologydevices.com/TMArrayer.htm  they have a manual arrayer that 
is running about 30,000.00. A little more than the Beecher Arrayer but it does 
offer some improvements. They are also able to sell the Needles that will fit 
into the Beecher instrument. I have spoken to Ron Gebing at Pathology Devices 
and he is responsive and reachable.

I am very optimistic, He has many units in use in the U.S. and the world.



Helen L. Fedor 

Tissue Microarray Lab, Manager
Prostate Spore Lab, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

410.614.1660





-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Barone, Carol 
Sent: Friday, March 04, 2011 3:56 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Beecher Instruments
Importance: High

Histonetters...It has been a longtime concern of mine that the only
company offering TMA instrumentation was Beecher Instruments.and the
wait list for those, was totally unacceptable. It came to my attention
today, from a reliable source at a university that Beecher has gone
out of business...( after buying Chemicon's arrayer...and taking away
the only real alternative). I have heard from  the same reliable source
they may be purchased by a foreign investor...my prayer is that perhaps
that could be Leica! Does anyone have more info on this? I wonder what
this means for those who already have one.at least I am on the wait
list!  We have tried several manual systems from molds to punchesand
there seems no real competition from these, which are difficult to
control or expensive to use.  What is the word out theresuggestions
for us "wait-ors". CB
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RE: [Histonet] Section position on slides

2011-02-16 Thread Helen Fedor
Since automation is becoming more and more a part of all Histology labs the 
demands of placement of the tissue on the slides varies for different 
instruments. Stainers, coverslippers and now with slide scanning as well. So I 
do not believe that there is a silver bullet answer.

Helen L. Fedor 

Tissue Microarray Lab, Manager
Prostate Spore Lab, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

410.614.1660


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bartlett, 
Jeanine (CDC/OID/NCEZID)
Sent: Wednesday, February 16, 2011 12:21 PM
To: Tanya Ewing-Finchem; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Section position on slides

I always try to center the section equally from sides and top-bottom as 
possible. This means measure from the bottom of the frosted edge as the "top".  
The Artisan special stains system has a clip that attaches around the slide to 
allow reagents to pool onto the sections and incubate.  If the section is too 
close to the sides then these areas do not stain adequately.  Sometimes if the 
tissue section itself is very large this is unavoidable.  With automated 
coverslippers you must also consider placement of tissue to allow for proper 
coverage.

If I am cutting multiple unstained slides for subsequent testing I try to 
orient the tissue the same on each slide to facilitate the reading of these 
slides by the pathologist's

Jeanine Bartlett
Infectious Diseases Pathology Branch
(404) 639-3590 
jeanine.bartl...@cdc.hhs.gov

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Tanya 
Ewing-Finchem
Sent: Wednesday, February 16, 2011 12:09 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Section position on slides


I am trying to put together a training document around microtomy and sectioning 
and am finding it hard to find information around the placement of the actual 
sections on the slides.  These are the objectives I am looking to answer.  Is 
this information found in any publications?
 
1)  Tissue / Section Placement:  Are there published guidelines / documentation 
on precisely where you should place tissue sections on a 25mm x 75mm glass 
slide?   Perhaps more importantly, where you should NOT place tissue (ie. "x" 
mm from the edge of the glass slide)?
 
2)  Diagnosable Slide Staining Area:  With automation becoming more widely used 
in IHC, are there published guidelines / documentation on the usable or 
diagnosable staining area on a 25mm x 75mm glass slide?  For instance, would 
you define that as the area under a traditional coverslip?  Would this be 
defined as the entire slide below the label?  Or is this some distance from all 
the edges of the slide?  With some automated systems, it is near impossible to 
get edge to edge staining.  Is this acceptable?  


 
Thanks for any ideas. 
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[Histonet] Charged slides

2011-01-06 Thread Helen Fedor
Hi, A question has come up regarding the different methods used to put a charge 
on the slide. We recently ordered some plus slides and the boxes they are 
packed in say silanized slides, but they say  "plus" on the slide . We don't 
want to use these for our clients if they are not going to be getting the same 
results as the former "plus" slides that we were using. I was under the 
delusion that Plus slides somehow are magically charged without any coating 
process taking place.

So does anyone know exactly how a "plus" slide gets its charge? Do they all get 
dipped in APES, or Polylysine?

Thanks for your help.

--
Helen

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RE: [Histonet] RE: Edge affect on TMA?

2010-12-22 Thread Helen Fedor
Here is a hint to keep your TMA from falling apart. After construction put the 
block face down on a glass slide a put in a 37degree C oven for 15-30 minutes 
(the paraffin should not be allowed to melt). Then take the block/slide out and 
gently compress with even pressure. (uneven pressure will cause the block to 
become misshapen). Cool the block at room temperature. You can repeat this 
process several times to help the cores adhere to the recipient block. The 
larger the core size, the more severe this problem is. One of the reasons that 
we choose not to use the 2.0 mm core is that this problem is more severe with 
this size core.

Hope that will help you.

Helen L. Fedor

Tissue Microarray Lab, Manager
Prostate Spore Lab, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

410.614.1660





From: IRENA SREBOTNIK KIRBIS [mailto:irena.kir...@hotmail.com]
Sent: Wednesday, December 22, 2010 1:54 AM
To: Helen Fedor; sette...@umdnj.edu; ahut...@dh.org; 
histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] RE: Edge affect on TMA?

how to prevent spots coming off during sectioning TM's? we do sometimes facing 
problems sectioning TM's, they're just falling apart?
thanks for any hints
Irena

> From: hfe...@jhmi.edu
> To: sette...@umdnj.edu; ahut...@dh.org; histonet@lists.utsouthwestern.edu
> Date: Tue, 21 Dec 2010 16:13:31 -0500
> CC:
> Subject: [Histonet] RE: Edge affect on TMA?
>
> Hello, We do not usually see that. If we do its very rare because I can't 
> think of any examples off hand. The usual problems are the spots came off or 
> shifted/smushed, not an edge effect.
>
> Helen
>
>
>
>
> -Original Message-
> From: histonet-boun...@lists.utsouthwestern.edu 
> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Settembre, 
> Dana
> Sent: Tuesday, December 21, 2010 3:00 PM
> To: 'Hutton, Allison'; histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Edge affect on TMA?
>
> Does anyone experience edge affect on TMA?
> Dana Settembre
>
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[Histonet] RE: Edge affect on TMA?

2010-12-21 Thread Helen Fedor
Hello, We do not usually see that. If we do its very rare because I can't think 
of any examples off hand. The usual problems are the spots came off or 
shifted/smushed, not an edge effect.

Helen




-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Settembre, Dana
Sent: Tuesday, December 21, 2010 3:00 PM
To: 'Hutton, Allison'; histonet@lists.utsouthwestern.edu
Subject: [Histonet] Edge affect on TMA?

Does anyone experience edge affect on TMA?
Dana Settembre

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RE: [Histonet] Pathology Grossing Manual

2010-12-09 Thread Helen Fedor
http://www.amazon.com/Surgical-Pathology-Dissection-Illustrated-ebook/dp/B000PY3QPM

here is the one that we use.

Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of tracz...@aol.com
Sent: Wednesday, December 08, 2010 6:34 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Pathology Grossing Manual


Amazon.com lists "Manual of Surgical Pathology Gross Room Procedures" by Juan 
Rosai.  Of course it's not available.  Does anyone know anything about this 
book?  Is it worth tracking down?  I need to update & expand an existing lab 
manual and thought it would be helpful.
Any ideas would be greatly appreciated.
Dorothy

MTA Histology LLC
PO Box 602 
Point Pleasant, NJ 08742
T:732-899-2912



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RE: [Histonet] unstained paraffin tissue slides storage--why cold?

2010-11-04 Thread Helen Fedor
Hello,  We have not published the paper yet . It is in progress and hope to get 
it out of the pile soon.

Helen

From: dusko trajkovic [mailto:dunat...@sbcglobal.net]
Sent: Thursday, November 04, 2010 1:45 PM
To: Morken, Tim; Helen Fedor
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] unstained paraffin tissue slides storage--why cold?

I did an experiment about 10 or 12 years ago, where I cut the sections and 
stained them at various intervals: Day1, Day 15, 1 month, 3 months, 6 months, 1 
year.
Before staining:
Set of slides were allowed to sit at room temp (RT).
Set of slides were stored at 4C.
Set of slides were vacuum sealed and stored at RT.
Set of slides were vacuum sealed and stored at 4C.

Slides stored at 4C consistently stained well, with a slight variance in 
staining after 1 year. Did not matter weather they were vacuum sealed or not.
Slides left out at RT did not fare so well. Staining variability was noticed 
after 1 month. When the slides were stained after one year, signal was almost 
eliminated. Variability and loss of antigenicity was observed with the vacuum 
sealed slides as well.
I did this experiment for just one antibody which was being extensively used at 
the time for one of our projects. It could be that other antibodies fare mach 
better under RT conditions, but why take a chance?

We keep all of our control slides at 4C.
Blocks are kept at RT.

Thanks
Dusko Trajkovic



From: "Morken, Tim" 
To: Helen Fedor 
Cc: "histonet@lists.utsouthwestern.edu" 
Sent: Thu, November 4, 2010 9:09:16 AM
Subject: RE: [Histonet] unstained paraffin tissue slides storage--why cold?

Helen, did you write a paper from that study?

Tim Morken
Supervisor, Histology, IPOX
UCSF Medical Center
San Francisco, CA, USA

-Original Message-
From: 
histonet-boun...@lists.utsouthwestern.edu<mailto:histonet-boun...@lists.utsouthwestern.edu>
 
[mailto:histonet-boun...@lists.utsouthwestern.edu<mailto:histonet-boun...@lists.utsouthwestern.edu>]
 On Behalf Of Helen Fedor
Sent: Thursday, November 04, 2010 8:56 AM
To: sgoe...@xbiotech.com<mailto:sgoe...@xbiotech.com>; Emily Sours
Cc: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>
Subject: RE: [Histonet] unstained paraffin tissue slides storage--why cold?

The study that we did showed that the staining on freshly cut slides from a 
block stored at room temperature was in fact not as good as slides that were 
sectioned 5 years earlier and stored at -20. Therefore the blocks should be 
stored in the cold as well. The tissue is degrading in the block and on the 
slides and the cold does slow down the process. The fixation in lots of the 
tissue is not optimal. Fixing for 48 hours will definitely  be better than the 
current fixation practices going on in most clinical labs,  if what you want is 
the best tissue preservation possible. But most of us do not have that option.

Helen L. Fedor

Tissue Microarray Lab, Manager
Prostate Spore Lab, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

-Original Message-
From: 
histonet-boun...@lists.utsouthwestern.edu<mailto:histonet-boun...@lists.utsouthwestern.edu>
 
[mailto:histonet-boun...@lists.utsouthwestern.edu<mailto:histonet-boun...@lists.utsouthwestern.edu>]
 On Behalf Of sgoe...@xbiotech.com<mailto:sgoe...@xbiotech.com>
Sent: Thursday, November 04, 2010 11:21 AM
To: Emily Sours
Cc: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>
Subject: RE: [Histonet] unstained paraffin tissue slides storage--why cold?

-70 or -80 seems a little extreme to me, that's why I always just leave
them in a normal freezer (-20).  I think the main point of doing this
from what I understand is so that the antigens stay "viable".  I know
over time they can degrade and so your stain won't work with some
antibodies.  The weirdest part to me has always been that you don't have
to store the blocks this way.  So I think that was your question, if the
blocks aren't stored in a freezer why store the slides?  Won't the
antigens in the blocks start to degrade as well?  This is a question I
would like to know the answer to as well...

Sarah Goebel, B.A., HT (ASCP)
Histotechnician


XBiotech USA Inc.

8201 East Riverside Dr. Bldg 4 Suite 100

Austin, Texas  78744

(512)386-2907




 Original Message 
Subject: Re: [Histonet] unstained paraffin tissue slides storage--why
cold?
From: Emily Sours mailto:talulahg...@gmail.com>>
Date: Thu, November 04, 2010 7:07 am
To: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>

Can I ask what the point of storing paraffin sections in freezing cold
storage?
They are wax sections, which never see any type of cold, so I don't
understand the point of this. I do understand putting them at 4 degrees
to
pre

RE: [Histonet] unstained paraffin tissue slides storage--why cold?

2010-11-04 Thread Helen Fedor
The study that we did showed that the staining on freshly cut slides from a 
block stored at room temperature was in fact not as good as slides that were 
sectioned 5 years earlier and stored at -20. Therefore the blocks should be 
stored in the cold as well. The tissue is degrading in the block and on the 
slides and the cold does slow down the process. The fixation in lots of the 
tissue is not optimal. Fixing for 48 hours will definitely  be better than the 
current fixation practices going on in most clinical labs,  if what you want is 
the best tissue preservation possible. But most of us do not have that option.

Helen L. Fedor 

Tissue Microarray Lab, Manager
Prostate Spore Lab, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of 
sgoe...@xbiotech.com
Sent: Thursday, November 04, 2010 11:21 AM
To: Emily Sours
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] unstained paraffin tissue slides storage--why cold?

-70 or -80 seems a little extreme to me, that's why I always just leave
them in a normal freezer (-20).  I think the main point of doing this
from what I understand is so that the antigens stay "viable".  I know
over time they can degrade and so your stain won't work with some
antibodies.  The weirdest part to me has always been that you don't have
to store the blocks this way.  So I think that was your question, if the
blocks aren't stored in a freezer why store the slides?  Won't the
antigens in the blocks start to degrade as well?  This is a question I
would like to know the answer to as well...

Sarah Goebel, B.A., HT (ASCP)
Histotechnician


XBiotech USA Inc.

8201 East Riverside Dr. Bldg 4 Suite 100

Austin, Texas  78744

(512)386-2907




 Original Message 
Subject: Re: [Histonet] unstained paraffin tissue slides storage--why
cold?
From: Emily Sours 
Date: Thu, November 04, 2010 7:07 am
To: histonet@lists.utsouthwestern.edu

Can I ask what the point of storing paraffin sections in freezing cold
storage?
They are wax sections, which never see any type of cold, so I don't
understand the point of this. I do understand putting them at 4 degrees
to
prevent mold, but -80 seems excessive.
We have kept our slides at room temperature for years and years, but
these
slides do not have an albumin coat (which I can see getting moldy), just
a
chemical coating.
Fixing for paraffin and paraffin infiltration seems to keep antigens
safe
without refrigeration because it's so intense, but that's just
conjecture on
my part.

Emily
--
Outside of a dog, a book is man's best friend. Inside of a dog it's too
dark
to read.
--Groucho Marx
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RE: [Histonet] unstained paraffin tissue slides storage

2010-11-03 Thread Helen Fedor
Hello, We have been storing our slides in very small Ziploc bags at -20 and 
find that this method works fairly well. We have done a study(Berez et.al in 
process) and slides stored in this fashion for 5 years stain better than 
freshly cut slides from the same blocks that have been stored at room 
temperature.


Helen L. Fedor 

Tissue Microarray Lab, Manager
Prostate Spore Lab, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

410.614.1660





-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of 
sgoe...@xbiotech.com
Sent: Wednesday, November 03, 2010 4:26 PM
To: Pop Elena
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] unstained paraffin tissue slides storage

Plain slide boxes is ok.  I think you can store them for up to a year in
a fridg. (4 degrees), but I usually pu them in a freezer (-20).

Sarah Goebel, B.A., HT (ASCP)
Histotechnician


XBiotech USA Inc.

8201 East Riverside Dr. Bldg 4 Suite 100

Austin, Texas  78744

(512)386-2907




 Original Message 
Subject: [Histonet] unstained paraffin tissue slides storage
From: Pop Elena 
Date: Wed, November 03, 2010 1:21 pm
To: histonet@lists.utsouthwestern.edu

Hello,
I found here a few disscussions regarding the storage of tissue slides
but I did 
not find a clear answer to the questions I have. I would really
appreciate an 
answer from anybody that has experience with this.

I need to store for long term a bunch of unstained tissue slides for the
purpose 
of doing immunostaining even in a few years from now on. Unfortunatelly
they 
were stored for about 3 years at room temperature. What it is 
usually recomended: to store them at -20 degrees Celsius? If yes, is it
OK to 
store them in the regular 100 slides boxes? And when you need to start
an 
immunostaining just take them out of the freezer and let them at room
temp for a 
while before starting the stain or what procedure do you use?

I heard some labs keep them in nitrogen gas containers. Do you have any
info 
about this?

Any imput is appreciated!
Thanks!



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RE: [Histonet] Block release

2010-10-18 Thread Helen Fedor
Hello, Our institution does release blocks, the requirement is that a patient 
consent is signed and we have an MTA (Material Transfer Agreement) For the 
study and appropriate IRB's in place for the study. It is a lot to set up, but 
after this is done for one study any following studies are easier to handle.

Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bernice 
Frederick
Sent: Monday, October 18, 2010 11:04 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Block release

Everyone,

I have been asked to post a query as to what your institution does in terms
of releasing blocks for oncology clinical trials. Please respond as it is
important to us as we receive a lot of those blocks here at Northwestern
(policies, procedures, alternative submissions etc) If you are in Australia,
Ireland, Canada, Puerto Rico or Peru please also answer (though I sort of
know already) as we do get blocks from you also.

Thanks

Bernice

 

 

Bernice Frederick HTL (ASCP)

Northwestern University

Pathology Core Facility

ECOGPCO-RL 

710 N Fairbanks Court

Olson 8-421

Chicago,IL 60611

312-503-3723

 

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RE: [Histonet] Cutting Microarrays

2010-09-20 Thread Helen Fedor
Hello, We have trouble with the larger size punches as well. In order to get 
the cores to stick to the size try warming the block in a oven at 37 degrees 
for 15 minutes, put the block face down on a clean slide. (providing the 
melting temperature of the paraffin is about 57 degrees). After 15 minutes 
remove the block/slide complex and gently press down on the block, then allow 
the block to cool. Repeat the process 3 or 4 times. We have also had some 
success by sliding the block face over the heating unit of the paraffin 
embedder. But this only works for a few sections and then you get the same 
problem back.

Best Regards,
Helen L. Fedor 

Tissue Microarray Lab, Manager
Prostate Tissue Spore Lab, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

410.614.1660





-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of louise renton
Sent: Monday, September 20, 2010 5:51 AM
To: Rae Staskiewicz; Histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Cutting Microarrays

hi, this sounds like the tissue was too cool when embedded in the wax - so
it has not made a nice homogenous block that cuts as one. Can u re-embed?

On Mon, Sep 20, 2010 at 10:54 AM, Rae Staskiewicz wrote:

>
>
> Good Morning All,
>
>
>
> I am having some issues when cutting a microarray containing 4mm punches.
> The punches are rolling and separating from the paraffin during cutting.
> Anyone have any tips or ideas??
>
>
>
> Rae Staskiewicz
>
> MMCI
>
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>



-- 
Louise Renton
Bone Research Unit
University of the Witwatersrand
Johannesburg
South Africa
+27 11 717 2298 (tel & fax)
073 5574456 (emergencies only)
"There are nights when the wolves are silent and only the moon howls".
George Carlin
No trees were killed in the sending of this message.
However, many electrons were terribly inconvenienced.
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RE: [Histonet] Cutting Microarrays

2010-09-20 Thread Helen Fedor
Hello, We have trouble with the larger size punches as well. In order to get 
the cores to stick to the size try warming the block in a oven at 37 degrees 
for 15 minutes, put the block face down on a clean slide. (providing the 
melting temperature of the paraffin is about 57 degrees). After 15 minutes 
remove the block/slide complex and gently press down on the block, then allow 
the block to cool. Repeat the process 3 or 4 times. We have also had some 
success by sliding the block face over the heating unit of the paraffin 
embedder. But this only works for a few sections and then you get the same 
problem back.

Best Regards,
Helen L. Fedor 

Tissue Microarray Lab, Manager
Prostate Tissue Spore Lab, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

410.614.1660

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of louise renton
Sent: Monday, September 20, 2010 5:51 AM
To: Rae Staskiewicz; Histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Cutting Microarrays

hi, this sounds like the tissue was too cool when embedded in the wax - so
it has not made a nice homogenous block that cuts as one. Can u re-embed?

On Mon, Sep 20, 2010 at 10:54 AM, Rae Staskiewicz wrote:

>
>
> Good Morning All,
>
>
>
> I am having some issues when cutting a microarray containing 4mm punches.
> The punches are rolling and separating from the paraffin during cutting.
> Anyone have any tips or ideas??
>
>
>
> Rae Staskiewicz
>
> MMCI
>
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>



-- 
Louise Renton
Bone Research Unit
University of the Witwatersrand
Johannesburg
South Africa
+27 11 717 2298 (tel & fax)
073 5574456 (emergencies only)
"There are nights when the wolves are silent and only the moon howls".
George Carlin
No trees were killed in the sending of this message.
However, many electrons were terribly inconvenienced.
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[Histonet] RE: Chilling trays

2010-07-01 Thread Helen Fedor
You can try disecting trays. I just googled discting tray, looks like listing 
price is 17.00. they are the perfect size and shape for freezing. and keep 
blocks organized.


Helen Fedor
Johns Hopkins University
TMA and Prostate Spore Lab Manager
JHU SOM Uro/Pathology
600 N. Wolfe Street, Marburg Room 406
Baltimore MD, 21287-7065
410-614-1660 / Fax 410-614-3695
Pager 410-283-3419



From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Breeden, Sara 
[sbree...@nmda.nmsu.edu]
Sent: Thursday, July 01, 2010 10:12 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Chilling trays

I went to eBay and found a set of three (total $12.00) stainless steel
"food prep" trays, roughly 9x5x2" (LxWxH). I fill them with DI water and
freeze them.   I also inherited a larger stainless steel container
that's 12.5x10.5x4.5" - same deal, DI water and freeze.   I face blocks,
put them on the ice and add a little DI water and soak for 10-15 minutes
- blocks cut like a dream! They stay frozen for a  l - o - n - g  time!
Try a local restaurant supply.



Sally Breeden, HT(ASCP)

Veterinary Diagnostic Services

New Mexico Department of Agriculture

700 Camino de Salud NE

Albuquerque, NM  87108

505-841-2576



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RE: [Histonet] RE: Prolonged FFPE slide and block storage

2010-05-15 Thread Helen Fedor
Hello, We have done a study and found that storage in small zip-loc bags at -20 
is an inexpensive and convenient way to store slides. In fact the slides stored 
for 5 years at -20 stain better than freshly cut sections from the same blocks. 
This was also published recently by others. Dr Rimm at Yale does have the 
storage under nitrogen gas system that is available commercially.


Helen L. Fedor 

Tissue Microarray Lab, Manager
Prostate Spore Lab, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

410.614.1660




-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Saturday, May 15, 2010 9:36 AM
To: SamuelPerry; histonet@lists.utsouthwestern.edu; TimMorken
Subject: Re: [Histonet] RE: Prolonged FFPE slide and block storage

Inexpensive long term anti oxidizing storage? Immerse them in mineral oil!
René J.

--- On Fri, 5/14/10, Morken, Tim  wrote:


From: Morken, Tim 
Subject: [Histonet] RE: Prolonged FFPE slide and block storage
To: "Perry, Samuel" , 
"histonet@lists.utsouthwestern.edu" 
Date: Friday, May 14, 2010, 5:05 PM


I have a paper from Yale several years ago about their tissue array facility. 
They found that storage in nitrogen helped a lot. I will try to find it, but 
you may be able to google it faster. 

Tim Morken
Supervisor, Histology / IPOX
UCSF Medical Center
San Francisco, CA  


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Perry, Samuel
Sent: Friday, May 14, 2010 1:16 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Prolonged FFPE slide and block storage

Hi All,
We have a growing need for storing slides. 

They need to be stored in a way that they won't become oxidized and loose their
antigenicity, since we use them for IHC.  

Any suggestions for inexpensive methods for prolonged storage of FFPE slides and
blocks is greatly appreciated.

Thanks!

Sam Perry

Research Technician 
Dana-Farber Cancer Institute
Boston, MA


The information in this e-mail is intended only for the person to whom it is
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[Histonet] Cover glass

2010-03-24 Thread Helen Fedor
Hello, Our lab does IHC staining in house and have had zero problems with our 
Fisher brand cover slips until recently. We've tried both Fisher and Corning 
cover slips and what appears to be dust or imperfections in the glass are found 
on both brands. This is creating false positives for us, because when the 
positive staining is low it is difficult to impossible to tell the 
imperfections from the staining itself.

Any advice will be welcome.

--
Helen

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[Histonet] RE: Akemi - Urgent!

2009-11-12 Thread Helen Fedor
I also receive one of the scam emails.


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Breeden, Sara
Sent: Thursday, November 12, 2009 9:21 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Akemi - Urgent!

Akemi Allison-Tacha - email me immediately, please.  I think you are
being scammed and I don't want to email you directly.

 

Sally Breeden, HT(ASCP)

NM Dept. of Agriculture

Veterinary Diagnostic Services

PO Box 4700

Albuquerque, NM  87106

505-841-2576

 

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RE: [Histonet] withdrawal

2009-10-21 Thread Helen Fedor
To subscribe or unsubscribe to the list or change your subscription to the 
daily digest mode etc. you need to go to:

http://lists.utsouthwestern.edu/mailman/listinfo/histonet

or email:

histonet-reque...@lists.utsouthwestern.edu with the word "help" (without 
quotation marks) in the subject line or body of the message. Instructions will 
be returned to you by email.






-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Roger Moretz
Sent: Wednesday, October 21, 2009 5:38 PM
To: ji...@bme.ogi.edu; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] withdrawal

The correct request is to "unsubscribe" but since you forgot the original 
information you received when you first subscribed or have failed to read any 
of the multitude of recent messages about unsubscribing, I'll guess you'll just 
have to suffer along with the rest of us in getting too many messages.

Roger Moretz, Ph.D.



- Original Message 
From: "ji...@bme.ogi.edu" 
To: histonet@lists.utsouthwestern.edu
Sent: Wed, October 21, 2009 4:26:58 PM
Subject: [Histonet] withdrawal

Hi

Please help me to withdrawal from the histonet.

I am frustrated to see a huge amount email each day.

Thanks
yali

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[Histonet] RE: re: control slide storage

2009-06-25 Thread Helen Fedor
Hello, We have a project that has worked on this topic using TMA's and have 
found that storing unstained, and unbaked slides at -20 enhances the 
preservation of antigenicity of the samples. It is not yet published but is 
getting written up. TMA slides that were stored for 5 years at -20 in very 
small Ziploc bags were compared to freshly cut TMA slides of the same block. 
Surprisingly some of the freezer slides tissue stained better then the freshly 
cut slides. So we need to start to store our tissue blocks in the cold as well.

--
Helen L. Fedor 

Tissue Microarray Lab, Manager
Prostate Spore Lab, Manager
Johns Hopkins University
600 N. Wolfe St, | Marburg Room 406
Baltimore, MD | 21287-7065

410.614.1660



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Renko, Heather 
D.
Sent: Thursday, June 25, 2009 4:44 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] re: control slide storage

Anyone have any  published literature for reference on storage control; 
refrigerator vs. room temp vs. freezer.  I have a friend asking for this and 
would also would love to get this information for my own reference  I searched 
the archives but, came up with only one publication.  Thank you in advance.

Heather D. Renko, Histology Supervisor
OSF Saint Anthony Medical Center
5666 East State Street
Rockford, Illinois 61108
Direct: 815-395-5410
heather.d.re...@osfhealthcare.org


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[Histonet] RE: Full Mount Immunos

2009-03-05 Thread Helen Fedor
I believe that Brain Research Labs has large "+" slides.
Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Metzger, Kenneth
Sent: Thursday, March 05, 2009 10:07 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Full Mount Immunos

Does anyone know of any companies producing charged full mount slides? I
have a pathologist that wants immunos done on full mount prostates and
the tissue keeps falling off. I have looked, but cannot find any charged
full mount slides. Any help would be appreciated!!

 

Kenneth G Metzger HTL(ASCP)

ARUP Labs

Histology Supervisor

500 Chipeta Way

Salt Lake City, Utah 84108-1221

kenneth.metz...@aruplab.com

(801) 583-2787 x 3101

 


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[Histonet] Pathos animal tissue processing

2009-02-19 Thread Helen Fedor
Hello, Another question, Is anyone using the Pathos system to process animal 
tissues? I would really appreciate any comments good/ and or bad on this topic.
Best regards,
Helen

From: Helen Fedor
Sent: Wednesday, February 18, 2009 5:33 PM
To: 'histonet@lists.utsouthwestern.edu'
Subject: Pathos Processing tissue problems

Hello, Our Surgical Pathology Department has been using the Pathos processor. I 
work in research and we have begun to have issues with the tissue falling off 
of the slide after pretreatment for IHC. Has anyone else noticed this 
happening? The tissue is much harder in the block than the standard method of 
processing. Is this normal? Does that have anything to do with the tissue 
falling off the slides?
Thanks in advance.


Helen L. Fedor

Comprehensive Tissue Services Center Manager
Prostate Spore Lab Manager
600 N. Wolfe St, Marburg Room 406
Baltimore, MD 21287-7065
410-614-1664
email:hfe...@jhmi.edu

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[Histonet] Pathos Processing tissue problems

2009-02-18 Thread Helen Fedor
Hello, Our Surgical Pathology Department has been using the Pathos processor. I 
work in research and we have begun to have issues with the tissue falling off 
of the slide after pretreatment for IHC. Has anyone else noticed this 
happening? The tissue is much harder in the block than the standard method of 
processing. Is this normal? Does that have anything to do with the tissue 
falling off the slides?
Thanks in advance.


Helen L. Fedor

Comprehensive Tissue Services Center Manager
Prostate Spore Lab Manager
600 N. Wolfe St, Marburg Room 406
Baltimore, MD 21287-7065
410-614-1664
email:hfe...@jhmi.edu

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RE: [Histonet] Cryostat safety question

2009-01-08 Thread Helen Fedor
Hello,  I had previously sent this but only to the person who asked the 
question, I think it this works beautifully so decided to resend to the list.

we have a great way to remove the specimens for the chucks. We have a 500cc 
plastic container with a lid that we have cut an "X" into the center.  We put 
warm water in the container and then just put the chuck stem into the "X". 
Within 10 seconds the block can be removed. Still frozen.  We just keep the 
plastic container at the bench and keep reusing the same one. The water level 
does need to come all the way up to the lid, But the chuck never gets wet. We 
do the sealing of the tissue with a tiny amount of OCT to the surface of the 
still frozen block, while it is still in the cryostat as well.


Helen

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Ingles Claire
Sent: Wednesday, January 07, 2009 7:38 PM
To: Histonet
Cc: mari.ann.mailh...@leica-microsystems.com
Subject: RE: [Histonet] Cryostat safety question

Andrea:
I work in a Mohs clinic where all we cut is frozen skin sections. Needless to 
say, we don't have 50 chucks laying around... In the morning before clinic 
starts we put a layer of freezing medium on chucks and put them in the cryostat 
to freeze. When we get specimens, we add another drop or so to the already 
frozen 'button' and immediately embed the tissue in it. We usually add another 
small drop on top after it has begun to freeze, to cover the specimen 
completely. Cut as normal when frozen. After done cutting all you have to do is 
use a forceps or other blunt object and pop the bit with the specimen in it 
away from the 'button' and return the chuck to the cryostat and it can be 
reused the rest of the day. The specimen is therefore still frozen for storage, 
and it has a quicker TAT. Plus you won't need nearly so many chucks, as they 
can be recycled almost as soon as you are done cutting. I usually keep 6-8 
'buttons' in my cryostat, and our clinic can process up to 50 separate 
specimens a day. A word of caution. If your work area is humid sometimes a thin 
layer of frost can form on the surface of the 'button' and when you attempt to 
take sections the bit with the tissue will pop off the 'button'. All you need 
to do is add another drop of medium to the button and 'glue' the two back 
together. If you are going a while between cutting sessions, I usually store my 
'buttons' upside(mountant side) down on one of the cryostat surfaces. It 
doesn't seem to develop the frost layer. Useful if you have tiny specimens.
Hope my verbose explanation is helpful. Feel free to e-mail if you have any 
questions or are confused about my explanation.

Claire Ingles
Madison WI



From: histonet-boun...@lists.utsouthwestern.edu on behalf of Andrea Hooper
Sent: Wed 1/7/2009 5:40 PM
To: Histonet
Cc: mari.ann.mailh...@leica-microsystems.com
Subject: [Histonet] Cryostat safety question



The discussion on microtome safety begs me to ask a cryostat question 

We have a Leica CM3050 cryostat and love it!

How are people (and perhaps only those in research do this) removing
their tissue from the chucks for future use? We often just section a
few slides worth then put the block at -80 deg C for future studies.
Needless to say, it's the most dangerous part of our day.

So what are your suggestions for removing tissue from a chuck (and
melting it isn't really a viable option)?

Thanks in advance,
Andrea
--

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