[Histonet] ANP.22780 and validation

[Cristi Rigazio via Histonet]

Hi Histoworld and Happy New Year!

I am currently validating new processing schedules and am a little concerned 
about my understanding.  NSH guidelines don't say special stains, IHCs etc., 
need to be re validated when validating a new processor, schedules etc., but 
this CAP standard seems to imply it is required.  This, in turn, doesn't make 
sense to me as many labs send their blocks or unstained slides for subsequent 
testing to other labs.  Those labs don't know the processing schedule, 
equipment, etc.,  Can someone shed some light on this for me?

Thank you,
Cristi

Sent from my iPhone
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Re: [Histonet] Fibroblasts

[Elizabeth Chlipala via Histonet]

Bernice

There are lots of different markers for fibroblasts via IHC - If you are 
looking for a good source Acris Antibodies has quite a few fibroblast markers

Vimentin, 
Smooth Muscle Actin
FSP-1 (fibroblast specific protein)
Pro Collagen I 
CD34
Prolyl-4-Hydroxylase beta
HSP-47

I have a two page document that I will send to you that lists a lot of what I 
have already and what types of fibroblasts that they stain for - we have used 
most of these in various species.

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

Ship to Address:

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


-Original Message-
From: Bernice Frederick via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Wednesday, January 04, 2017 1:02 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Fibroblasts

Hello all,
Don't have a ton of time to search so am asking. Any good markers specific to 
fibroblasts? And am drawing a blank for regular special stains for some reason. 
Researcher says Vimentin is not specific 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] Grad Student Problem

[Elizabeth Chlipala via Histonet]

You can also thaw and refreeze we have done that before.  

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

Ship to Address:

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


-Original Message-
From: Terri Braud via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Wednesday, January 04, 2017 12:16 PM
To: 'histonet@lists.utsouthwestern.edu'
Cc: 'r...@psu.edu'
Subject: Re: [Histonet] Grad Student Problem

My response:
My first question is what temperature are the blocks that are too brittle? And 
at what temperature are you trying to cut?  Blocks stored in cryofreezers at 
-70o C or less are far too cold to cut without brittleness.  My suggestion 
would be to pull the blocks and put them into the cryostat at -18o C for at 
least 6 hours to acclimate to that temp, then try to cut and stain for IF.

If that doesn't work, then I would thaw in formalin and process as routine 
tissue for formalin fixed paraffin embedded.  The process that your student 
described in the 2nd step is not a complete, or valid process.
Depending on the T-Cell markers they are trying to demonstrate, one can 
successfully use standard IHC with appropriate clones, or with usually less 
success, use a modified IF stain procedure for FFPE sections. Since there are a 
"ba-zillion" T-cell markers, any further details are very dependent on the 
markers and the condition of the tissue.
Sincerely, Terri Braud

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

   1. grad student problem (Roberta Horner)
Message: 1
Date: Tue, 3 Jan 2017 18:05:15 +
From: Roberta Horner 
I got the following from a grad student here at Penn State. I am not sure how 
to solve his problem if possible. Does anyone have any suggestions I can 
forward?
Roberta Horner
Animal Diagnostic Lab
Penn State University

"I am having some difficulties sectioning mouse tumor samples for 
immunofluorescent analysis.  We originally went the OCT route because we are 
staining for T cell markers and were worried that the heating that occurs 
during paraffin embedding would compromise the T cell receptor.  The samples 
are a little old, but we are hoping to section and stain for immune cell 
infiltrates.  When sectioning with the cryostat, the tissue and OCT is quite 
brittle and the sample is not intact enough to transfer to a slide. Two 
colleagues have given the following suggestions:
1. Thaw the OCT blocks, remove the tissue, and place in a new block with fresh 
OCT media.  I've tried this technique on a few practice samples and the new OCT 
media seems to be less brittle and I'm able to get tissue on a slide, however 
the tissue itself still seems to be poor with either freeze or sectioning 
artifacts.
2. Thaw the OCT blocks in water, remove the tissue and place in formalin 
overnight, place in 70% EtOH, then paraffin embed.  Section on a microtome.  
Check the fluorescently labelled antibody data sheet to see if paraffin 
embedding interferes with binding.  Try to stain and see what happens.

I was hesitant to try the second suggestion because I have found no protocol 
that takes tissue originally stored in OCT blocks and subsequently redirects 
them to formalin and paraffin for microtome sectioning.  If you have any 
recommendations on how to move forward and section these difficult samples, or 
know anyone at the diagnostics lab or at Penn State that could help, that would 
be much appreciated!"

*


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[Histonet] Histology school in NJ

[Ann Specian via Histonet]

Does anyone know where to find the requirements to start aschool for 
Histotechnology in nj?


Ann
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[Histonet] Fibroblasts

[Bernice Frederick via Histonet]

Hello all,
Don't have a ton of time to search so am asking. Any good markers specific to 
fibroblasts? And am drawing a blank for regular special stains for some reason. 
Researcher says Vimentin is not specific 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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[Histonet] Questions regarding microwave questions on Anatomic Pathology CAP Checklist

[Piche, Jessica via Histonet]

Happy New Year Everyone!!  :)

Questions on CAP Anatomic Path Checklist. Are the microwave questions referring 
to microwave processors/processing or the microwave oven that we use for 
special stains? Questions are ANP. 27170, 28290, 28860, and 29430.

I'm thinking the microwave oven but I want to confirm or deny.

Thanks in advance,

Jessica Piche, HT(ASCP)
Waterbury Hospital



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Re: [Histonet] Grad Student Problem

[Terri Braud via Histonet]

My response:
My first question is what temperature are the blocks that are too brittle? And 
at what temperature are you trying to cut?  Blocks stored in cryofreezers at 
-70o C or less are far too cold to cut without brittleness.  My suggestion 
would be to pull the blocks and put them into the cryostat at -18o C for at 
least 6 hours to acclimate to that temp, then try to cut and stain for IF.

If that doesn't work, then I would thaw in formalin and process as routine 
tissue for formalin fixed paraffin embedded.  The process that your student 
described in the 2nd step is not a complete, or valid process.
Depending on the T-Cell markers they are trying to demonstrate, one can 
successfully use standard IHC with appropriate clones, or with usually less 
success, use a modified IF stain procedure for FFPE sections. Since there are a 
"ba-zillion" T-cell markers, any further details are very dependent on the 
markers and the condition of the tissue.
Sincerely, Terri Braud

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

   1. grad student problem (Roberta Horner)
Message: 1
Date: Tue, 3 Jan 2017 18:05:15 +
From: Roberta Horner 
I got the following from a grad student here at Penn State. I am not sure how 
to solve his problem if possible. Does anyone have any suggestions I can 
forward?
Roberta Horner
Animal Diagnostic Lab
Penn State University

"I am having some difficulties sectioning mouse tumor samples for 
immunofluorescent analysis.  We originally went the OCT route because we are 
staining for T cell markers and were worried that the heating that occurs 
during paraffin embedding would compromise the T cell receptor.  The samples 
are a little old, but we are hoping to section and stain for immune cell 
infiltrates.  When sectioning with the cryostat, the tissue and OCT is quite 
brittle and the sample is not intact enough to transfer to a slide. Two 
colleagues have given the following suggestions:
1. Thaw the OCT blocks, remove the tissue, and place in a new block with fresh 
OCT media.  I've tried this technique on a few practice samples and the new OCT 
media seems to be less brittle and I'm able to get tissue on a slide, however 
the tissue itself still seems to be poor with either freeze or sectioning 
artifacts.
2. Thaw the OCT blocks in water, remove the tissue and place in formalin 
overnight, place in 70% EtOH, then paraffin embed.  Section on a microtome.  
Check the fluorescently labelled antibody data sheet to see if paraffin 
embedding interferes with binding.  Try to stain and see what happens.

I was hesitant to try the second suggestion because I have found no protocol 
that takes tissue originally stored in OCT blocks and subsequently redirects 
them to formalin and paraffin for microtome sectioning.  If you have any 
recommendations on how to move forward and section these difficult samples, or 
know anyone at the diagnostics lab or at Penn State that could help, that would 
be much appreciated!"

*


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[Histonet] Greenbay Histology Opportunities with growing lab

[Angie Laparidis via Histonet]

Happy New Year HistoNet Users!


I send out an email last year with news of an expanding client in the
*Greenbay,
WI* area. They successfully filled their opening, and have some good news
for the new yearthey are continuing to grow!!

This private lab has a new opening on their 2nd shift as well as a new
opening on their 3rd shift. Their compensation and benefits packages are
unmatched and they truly take care of their employees!


Let me know if you can think of anyone who might be a good fit:)


Sincerely,


*Angie Laparidis*Healthcare Recruiter
K.A. Recruiting, Inc.
10 Post Office Square, 8th Floor South, Boston, MA, 02109
W:  617.746-2744 (*please note this is a new number*)
F: (617) 507-8009
an...@ka-recruiting.com


Our openings are updated daily at www.ka-recruiting.com
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[Histonet] p16 replacements?

[Cassie P. Davis via Histonet]

Hi Histonet Folks,

our current vendor  will be discontinuing carrying the antibody for 
p16. Does anybody have suggestions for replacing in it? We currently use it to 
stain for CIN. Many thanks in advance for sharing.


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



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[Histonet] Beecher (Estigen) ATA27 tissue arrayer for sale

[Donato, Elizabeth M via Histonet]

Hello Everyone,

We are selling our Beecher (now Estigen) ATA 27 automated tissue arrayer and 
before I post it on re-sale websites, I'd like to share this information with 
the Histonet community.

This instrument was completely re-built/refurbished in 2012 and I have all 
supporting documentation.  It's essentially a brand new instrument.  TMA blocks 
can be constructed using cores of various diameters ranging in size from 0.6mm 
to 3.0 mm.  Cores (0.6mm) can be transferred at a rate of approximately 120-180 
cores per hours and up to 26 replicate TMA blocks can be constructed during a 
single run. This robotic unit can be fully controlled via a high-performance PC 
pre-loaded with custom software for mapping donor cores to a single or multiple 
recipient blocks.  It has an external video camera which is an upgraded 
feature.  Using this TMA construction software this external camera allows you 
to superimpose the image of a target marked H over the image of the 
corresponding paraffin block surface for accurate donor core targeting.  The 
user interface is straightforward and gives the you complete control over 
optimizing variables to accommodate variations in tissue characteristics b
 etween donor blocks.  Once the TMA grid has been created and the donor blocks 
mapped using the software, no further user interaction is needed.

The nature of our TMA projects has changed over the last few years and this 
instrument is now underutilized.  It needs to be in a lab where it can be used 
at full capacity.  I have a recent quote for a new ATA27 at a cost of 180K, so 
if you are looking to purchase high throughput TMA construction equipment, this 
instrument could save you a significant amount of money.  We are still in the 
process of establishing a selling price.

If you'd like additional information, pictures or a demonstration of this 
instrument's capabilities, please contact me.

Best,

Liz

Elizabeth Donato MT(ASCP)
Manager | Porter and Specialized Pathology Laboratories
*
206.667.4501 p | 206.667.5815 f | 
edon...@fredhutch.org

Fred Hutchinson Cancer Research Center
1100 Fairview Ave. N., Mail Stop C1-015
Seattle, WA 98109
fredhutch.org





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