[Histonet] Re: Histonet Digest, Vol 101, Issue 27

[Amos Brooks]

Hi,
 Strange issue with your Ki-67. I don't know what would do that unless
it was non-specific background. Is it nuclear or cutoplasmic?
 If you are working in mice or other animals you could try labeling
them with BRDu then detecting it with an anti BRDu antibody. Humans don't
like being injected with BRDu then being sacrifices but lawyers do. Also
Cleaved Caspase 3 works nicely and is probably more specific to apoptosis
than Tunel which also picks up necrosis.

Have a great weekend,
Amos


On Fri, Apr 20, 2012 at 1:00 PM,
wrote:

> Message: 9
> Date: Thu, 19 Apr 2012 15:07:43 -0500
> From: "Margaryan, Naira" 
> Subject: [Histonet] proliferation and apoptosis
> To: "histonet-requ...@lists.utsouthwestern.edu"
>
> Cc: "histonet@lists.utsouthwestern.edu"
>
> Message-ID:
><
> c1ba93040c6b9a4a8d84878f93fec36a0ab7d0e...@cmhexcc01mbx.childrensmemorial.org
> >
>
> Content-Type: text/plain; charset="us-ascii"
>
> Hi histonetters,
>
> I am looking for the good markers to detect (separately) proliferation and
> apoptosis of cells in tumor sections. Unfortunately, KI-67 stains apoptotic
> bodies as well as proliferated cells; and the tunnel assay shows both
> apoptotic body and proliferation.
>
> Any suggestions for the FFPE tissue and the HRP protocol are appreciated.
>
> Thanks in advance,
> Naira
>
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Re: [Histonet] Leica Peloris Processor

[Patrick Laurie]

We have 4 peloris processors, run many IHC and FISH stains, none of them
have had any negative effects due to the increased temperatures.  The
increased temps aren't that high, 45 degrees for all the liquids (we use
xylene) and 65 degrees for the paraffins.

Good luck

Patrick

On Fri, Apr 20, 2012 at 12:47 PM, Joe W. Walker, Jr. wrote:

> Hello all,
>
> I am new to your listserve.  I am wondering if anyone has experience with
> the Leica Peloris processor.  Specifically, has anyone noticed if the
> higher processing temperatures has affected any IHC staining results or
> FISH results.
>
> Looking forward to responses,
>
> Joe W. Walker, Jr. SCT(ASCP)CM
> Anatomical Pathology Manager
> Rutland Regional Medical Center
> 160 Allen Street, Rutland, VT 05701
> Phone: 802.747.1790  Fax:802.747.6525
> NEW EMAIL: joewal...@rrmc.org
> www.rrmc.org
>
> Our Vision: To be the Best Healthcare System in New England
> Rutland Regional...Vermont's 1st Hospital to Achieve Both ANCC Magnet
> Recognition(r) and the Governor's Award for Performance Excellence
>
> This message (and any included attachments) is from Rutland Regional
> Health Services and is intended only for the addressee(s). The information
> contained herein may include privileged or otherwise confidential
> information. Unauthorized review, forwarding, printing, copying,
> distributing, or using such information is strictly prohibited and may be
> unlawful. If you received this message in error, or have reason to believe
> you are not authorized to receive it, please promptly delete this message
> and notify the sender by e-mail.
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-- 
Patrick Laurie HT(ASCP)QIHC
CellNetix Pathology & Laboratories
1124 Columbia Street, Suite 200
Seattle, WA 98104
plau...@cellnetix.com
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Re: [Histonet] Leica Peloris Processor

[Richard Cartun]

In my experience, no.  Our current IHC/ISH is as good, if not better than what 
we were getting with our former processors.

Richard

Richard W. Cartun, MS, PhD
Director, Histology & Immunopathology
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596 Office
(860) 545-2204 Fax


>>> "Joe W. Walker, Jr."  4/20/2012 3:47 PM >>>
Hello all,

I am new to your listserve.  I am wondering if anyone has experience with the 
Leica Peloris processor.  Specifically, has anyone noticed if the higher 
processing temperatures has affected any IHC staining results or FISH results.

Looking forward to responses,

Joe W. Walker, Jr. SCT(ASCP)CM
Anatomical Pathology Manager
Rutland Regional Medical Center
160 Allen Street, Rutland, VT 05701
Phone: 802.747.1790  Fax:802.747.6525
NEW EMAIL: joewal...@rrmc.org
www.rrmc.org

Our Vision: To be the Best Healthcare System in New England
Rutland Regional...Vermont's 1st Hospital to Achieve Both ANCC Magnet 
Recognition(r) and the Governor's Award for Performance Excellence

This message (and any included attachments) is from Rutland Regional Health 
Services and is intended only for the addressee(s). The information contained 
herein may include privileged or otherwise confidential information. 
Unauthorized review, forwarding, printing, copying, distributing, or using such 
information is strictly prohibited and may be unlawful. If you received this 
message in error, or have reason to believe you are not authorized to receive 
it, please promptly delete this message and notify the sender by e-mail.

Thank You
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[Histonet] Leica Peloris Processor

[Joe W. Walker, Jr.]

Hello all,

I am new to your listserve.  I am wondering if anyone has experience with the 
Leica Peloris processor.  Specifically, has anyone noticed if the higher 
processing temperatures has affected any IHC staining results or FISH results.

Looking forward to responses,

Joe W. Walker, Jr. SCT(ASCP)CM
Anatomical Pathology Manager
Rutland Regional Medical Center
160 Allen Street, Rutland, VT 05701
Phone: 802.747.1790  Fax:802.747.6525
NEW EMAIL: joewal...@rrmc.org
www.rrmc.org

Our Vision: To be the Best Healthcare System in New England
Rutland Regional...Vermont's 1st Hospital to Achieve Both ANCC Magnet 
Recognition(r) and the Governor's Award for Performance Excellence

This message (and any included attachments) is from Rutland Regional Health 
Services and is intended only for the addressee(s). The information contained 
herein may include privileged or otherwise confidential information. 
Unauthorized review, forwarding, printing, copying, distributing, or using such 
information is strictly prohibited and may be unlawful. If you received this 
message in error, or have reason to believe you are not authorized to receive 
it, please promptly delete this message and notify the sender by e-mail.

Thank You
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RE: [Histonet] GMS on Toenail

[Britton, Josette C]

The way we got around that problem was to do a manual PAS stain instead!


Josie Britton HT
Cheshire Medical Center
Keene NH

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Scott,
Allison D
Sent: Thursday, April 19, 2012 3:23 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] GMS on Toenail

Hello to all in histoland.  We have a stubborn toenail that keeps coming
off when we try to do a GMS stain on the ventana machine.  Any
suggestions on how to keep the section on the slide during the staining
procedure.

Allison Scott HT(ASCP)
Histology Supervisor
LBJ Hospital
Houston, Texas

CONFIDENTIALITY NOTICE:
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[Histonet] Immunofluorescence

[Thotakura, Anil Kumar]

Dear All,

Can some one please send the protocol for immunofluorescence, I want to stain 
for FITC CD45.1 on mice spleens.

Thank you very much for your help.

Many Thanks,
Anil kumar.
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[Histonet] Re: Pinning Specimens

[Bob Richmond]

About pinning specimens so they fix flat: Margaret Horne notes the use
of dental wax. I've used it to make small "boats" for the rather
exacting procedure of pinning muscle biopsy specimens for electron
microscopy. Here a 1 to 2 mm bundle of longitudinal fibers has to be
gently stretched to its resting length, pinned in the "boat", and
promptly fixed in glutaraldehyde.

One technologist noted that she had a young pathologist who was "wet
behind the ears" and wanted to pin specimens, and was looking to the
older pathologists to bring him in line to The Way We've Always Done
It. I try to remain "wet behind the ears", though after nearly 50
years of occasionally pinning specimens I find myself a little dry.

We continue to lose skills in the gross room. Today I'm working with a
service that has almost no ability to handle calcified tissue - I need
to go to the hardware store and buy a hacksaw to replace the worn-out
Satterlee saw, which I think was gathered up off a Civil War
battlefield.

Bob Richmond
Samurai Pathologist
Knoxville TN

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[Histonet] Re: Histonet Digest, Vol 101, Issue 27

[Eric Tambutte]

Bonjour,
Je suis absent du laboratoire jusqu'au jeudi 03 mai 2012. Je vous répondrai le 
plus rapidement possible.

Eric Tambutté

Thank you for your mail. I will be out of office till May 03rd 2012.
I will respond to your e-mail as soon as possible.  
Thank you for your understanding.
Best regards
Eric Tambutté

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[Histonet] Part Time Job Opening, Springfield, MA

[Amanda L]

Please Send Resume for: 

Part Time opportunity for a Histotechnican. 

Duties and responsibilities include:
• Under general supervision performs routine and non-routine activities 
involved in the preparation of slides, for microscopic evaluation by 
pathologist(s) according to policies and procedures
• Process paperwork associated with accessioning and reporting. 
• Eligible to gross per CLIA requirements.
• Ensure proper tissue processing. 
• Embed processed tissue in paraffin
• Prepare slides for routine staining. 
• Perform microtomy of embedded tissue
• Perform coverslipping of stained slides either manually or automated. 
• Perform filing of finished blocks and slides. 
• Perform routine maintenance and cleaning of equipment and troubleshoot minor 
equipment failures. Document remedial actions such as repairs or repeated 
tests. 
• Adhere to laboratory's quality control policies, and document all quality 
control activities. 
• Ensure all corporate safety, quality control and quality assurance standards 
are met. 
• Ensure compliance with all local, federal, CLIA and CAP regulations. 
• Maintain a clean and well-organized work area. 
• Clinical Chemistry & other duties, as assigned by supervisor. 

Minimum Requirements Include: 
• AA or AS degree or equivalent training and experience , CLIA approved for 
grossing specimens
• HT (ASCP) or HTL (ASCP) eligible OR five years of full time experience in the 
last 10 years. 
• Work Experience: 1-3 years experience as a Histotechnician in an anatomic 
pathology laboratory.
• License for Clinical Laboratory Technician or Technologist license. 
***Note this is a part time position also with coverage for vacation 

Thanks!   
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[Histonet] Re: Pinning Specimen

[Margaret Horne]

Just another idea : I have used dental wax. Comes in a package of about
20 sheets pf pink wax, each 7.5 cm x 14.5 cm x 0.2 cm. It's a little bit
more pliable than the  waste paraffin, but not as cheap :-)  I cut it in
a long rectangle so that it sticks out of the formalin  with the tissue
at the bottom end , rather like  celery in a Bloody Caesar.  
Margaret
 

>>> Bob Richmond  19/04/2012 4:44 PM >>>
Karen Heckford HT ASCP CE at St. Mary's Medical Center in San Francisco
asks:

>>Does anyone know where to get specimen boards that you can pin
specimens to and then submerse in formalin? I ordered them a long time
ago and cannot remember where I got them.<<

I've solved this problem several ways. Probably the best, which has
already been mentioned, is to have blocks cast from waste paraffin, in
several sizes, and pin the specimen to them with those T-shaped steel
"map pins" and put it face down in the fixative. You can also use
cardboard or styrofoam. You may need to put a weight on top of the
pinning board. Most specimens need fixing overnight. You can ink
before or after fixation.

Most of the pathology services I've worked in do not have such pinning
arrangements and do not welcome them. Does anyone know what the
Hospital Administrator and Lab Manager's Handy-Dandy Manual for Tying
the Pathologist in Knots has to say about them? (The top-secret book
they all have in their desk drawers.)

Bob Richmond
Samurai Pathologist
Knoxville TN

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[Histonet] re: Pin 4

[Robert Jann]

Karen,

Here's our protocol for FFPE sections on the Benchmark XT.  Be advised that the 
times will be approximate for you and that we use Dako P504s @ 1:50.  Also, you 
can call Ventana and request a rep. to come to your site and help you optimize 
the stain.

1.  Create/manage protocols
2.  select IHC DS uDAB - uRed v3
3.  select deparaffinization
4.  select cell conditioning - mild CC1
5.  select antibody - 34BE12 + p63 (4536) for 8 min.
6.  select ultrawash
7.  select antibody denaturization - 95 deg. C for 4 min.
8.  select DS antibody - Prep kit 1 (2851) (P504s) for 48 min.
9.  select DS ultrawash
10.  select counter stain - hematoxylin for 4 min. and blue for 4 min.

You will also need to use the ultraview universal DAB detection kit and ultra 
view universal alkaline phosphatase red detection kit.  Good luck in setting it 
up.

Bob Jann HT(ASCP)
 
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