Re: [Histonet] Question Regarding HM355S Automatic Microtome

[Donna Emge via Histonet]

Diana,

I have used the HM355S at two different labs and liked it. Make sure to
demo it or purchase it with the “E” type disposable blade carrier. No one
liked the “ER” type blade carrier.  You can see the difference between them
in the operator manual -?just do a browser search for the manual. I also
really liked using the foot pedal to stop and start. The memory feature
makes fast work of. trimming blocks. When you put the next block in the
clamp, push the memory button and it retracts to the exact position needed
to start trimming the block.

Donna Emge
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Re: [Histonet] Question Regarding HM355S Automatic Microtome

[Robyn L Vazquez via Histonet]

Hello,
I have been using this microtome for the past 12 years. Love it! Regular 
maintenances and it will last a longtime. Just changed the motherboard 
(expensive) and back in business. Can not thing of any cons all good.

Robyn

-Original Message-
From: Piche, Jessica via Histonet 
Sent: Thursday, February 29, 2024 3:50 AM
To: histonet@lists.utsouthwestern.edu; Diana Martinez-Longoria 

Subject: Re: [Histonet] Question Regarding HM355S Automatic Microtome

Caution: This email came from outside Kaiser Permanente. Do not open 
attachments or click on links if you do not recognize the sender.

__
Hi Diana,

We are not currently using the HM355S in our lab, but we did demo it and were 
not fans. I would see if you can demo one to see if you like it.  We had the 
Thermo Shandon Finesse and Finesse Me models and we loved them but the HM355S 
didn't appear to be designed the same way and we ended up getting the Leica 
Autocuts and love them!!

Have a great day.

Jessica

Jessica Piche, HT(ASCP)
Waterbury Hospital Histology Laboratory
Histology Team Leader
203-573-7167

From: Diana Martinez-Longoria via Histonet 
Sent: Wednesday, February 28, 2024 4:46 PM
To: histonet@lists.utsouthwestern.edu 
Subject: [Histonet] Question Regarding HM355S Automatic Microtome

[EXTERNAL MSG]

Hello All,

I am reaching out to seek help regarding if anyone is currently or have in the 
past used the HM355S Automatic Microtome? What are the pros and cons?
Thank you in Advance!
Diana Martinez-Longoria
El Centro Regional Medical Center
Lead Histotechnician (ASCP)cm
Laboratory - Pathology Department
1415 Ross Ave | El Centro, CA  92243
760.339.7267: Fax: 760-3394570
 diana.martinez-longo...@ecrmc.org<mailto:diana.martinez-longo...@ecrmc.org>






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Re: [Histonet] Question Regarding HM355S Automatic Microtome

[Piche, Jessica via Histonet]

Hi Diana,

We are not currently using the HM355S in our lab, but we did demo it and were 
not fans. I would see if you can demo one to see if you like it.  We had the 
Thermo Shandon Finesse and Finesse Me models and we loved them but the HM355S 
didn't appear to be designed the same way and we ended up getting the Leica 
Autocuts and love them!!

Have a great day.

Jessica

Jessica Piche, HT(ASCP)
Waterbury Hospital Histology Laboratory
Histology Team Leader
203-573-7167

From: Diana Martinez-Longoria via Histonet 
Sent: Wednesday, February 28, 2024 4:46 PM
To: histonet@lists.utsouthwestern.edu 
Subject: [Histonet] Question Regarding HM355S Automatic Microtome

[EXTERNAL MSG]

Hello All,

I am reaching out to seek help regarding if anyone is currently or have in the 
past used the HM355S Automatic Microtome? What are the pros and cons?
Thank you in Advance!
Diana Martinez-Longoria
El Centro Regional Medical Center
Lead Histotechnician (ASCP)cm
Laboratory - Pathology Department
1415 Ross Ave | El Centro, CA  92243
760.339.7267: Fax: 760-3394570
 diana.martinez-longo...@ecrmc.org<mailto:diana.martinez-longo...@ecrmc.org>






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[Histonet] Question Regarding HM355S Automatic Microtome

[Diana Martinez-Longoria via Histonet]

Hello All,

I am reaching out to seek help regarding if anyone is currently or have in the 
past used the HM355S Automatic Microtome? What are the pros and cons?
Thank you in Advance!
Diana Martinez-Longoria
El Centro Regional Medical Center
Lead Histotechnician (ASCP)cm
Laboratory - Pathology Department
1415 Ross Ave | El Centro, CA  92243
760.339.7267: Fax: 760-3394570
 diana.martinez-longo...@ecrmc.org






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e-mail and its attachments. 
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Re: [Histonet] Question about 72 hour upper fixation time for HER2

[Whitaker, Bonnie via Histonet]

Well, I would worry much less about over-fixation than I would under-fixed 
tissues.  Do you not have the option to have a weekend processing schedule that 
runs RT formalin?

Is your volume sufficient you could do an analysis of weekend vs non-weekend 
ER, PR, HER2 rates and see if they are statistically different?  You should be 
keeping the overall statistics already.  If they are not statistically 
different, that should ease your mind,

Good luck!
Bonnie Whitaker




Get Outlook for iOS<https://aka.ms/o0ukef>

From: Tony Auge via Histonet 
Sent: Friday, May 5, 2023 6:50:00 PM
To: histonet@lists.utsouthwestern.edu 
Subject: [Histonet] Question about 72 hour upper fixation time for HER2

Hi Histonet,

Does anyone know if the 72 hour upper fixation time for HER2 was calculated
at room temperature? My lab is currently running our formalin at 40°c and
we go 72 hours on the long weekends, so I'm worried it might be overfixed.

Thanks,
Tony

--

Tony Auge HTL (ASCP)CM QIHC
Cell: (651) 373-4768
Email: tony.a...@gmail.com
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[Histonet] Question about 72 hour upper fixation time for HER2

[Tony Auge via Histonet]

Hi Histonet,

Does anyone know if the 72 hour upper fixation time for HER2 was calculated
at room temperature? My lab is currently running our formalin at 40°c and
we go 72 hours on the long weekends, so I'm worried it might be overfixed.

Thanks,
Tony

-- 

Tony Auge HTL (ASCP)CM QIHC
Cell: (651) 373-4768
Email: tony.a...@gmail.com
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Re: [Histonet] Question regarding posting job openings

[John Shelton via Histonet]

Howdy Lisa and Histonet List Members,

It is permitted. I think the membership benefits from knowing what 
opportunities exist in the field.
Job postings via recruitment services, as well as those from individual 
hospitals, commercial, or academic labs are welcome.

Logistics on how to accomplish...
The poster would need to be a subscriber to Histonet.
The message would need to be 60kb or smaller, so I suggest stripping any 
signature logos from the post.

Good luck in your search for the right candidate.

JS



John M Shelton
Core Operations Manager
Histonet List Owner & Moderator

UT Southwestern
Histo Pathology Core
5323 Harry Hines Blvd
Dallas, Texas 75390-8573
(214)648-1451

Visit the Core's website at:
http://www.utsouthwestern.edu/labs/histo-pathology/

Visit Histonet Archives at:
http://www.histosearch.com/histonet.html




From: zzz_Tag_histonet-lists 
Sent: Thursday, April 21, 2022 7:41 AM
To: zzz_Tag_histonet-lists 
Subject: [Histonet] Question regarding posting job openings

Is it permitted for a company to post job openings in Histonet? If so, how does 
that company go about posting it?

Lisa Freeman, AAS, HT
Histology Supervisor
Toxicologic Pathology Associates
National Center for Toxicological Research
3900 NCTR RD
Jefferson, AR 72079
Phone: 870-543-7234
E-mail: lisa.free...@fda.hhs.gov<mailto:lisa.free...@fda.hhs.gov>

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UT Southwestern

Medical Center

The future of medicine, today.
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[Histonet] Question regarding posting job openings

[Freeman, Lisa* via Histonet]

Is it permitted for a company to post job openings in Histonet? If so, how does 
that company go about posting it?

Lisa Freeman, AAS, HT
Histology Supervisor
Toxicologic Pathology Associates
National Center for Toxicological Research
3900 NCTR RD
Jefferson, AR 72079
Phone: 870-543-7234
E-mail: lisa.free...@fda.hhs.gov

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[Histonet] Question for Research on Bone

[Porter, Amy via Histonet]

So I am working with Canine femurs and tibias that have been dissected as 
medial and lateral - they have been decaled in formic acid - rinsed in running 
tap well - processed on a routine 1 hour processing program . cutting very 
well!!

My issue is that the articular cartilage on the bone surface keep flipping up 
and over onto itself - I am using a 37C slide warmer overnight to allow them to 
hopefully flatten out more than on the water bath and then drying in the 56C 
oven for about 1 hour - then staining on automated Leica stainer - they are 
intact at 37. look like the might be curling at 56C??

Anyone have any suggestions???

TIA - Amy

Amy S. Porter, HT (ASCP) - Lab Supervisor
Michigan State University
Investigative HistoPathology Lab
567 Wilson Road - Rm 2133
East Lansing, MI  48824
Ph:  517-884-5026
(she/her/hers)
https://sites.google.com/msu.edu/ihpl/home
"Life is like riding a bicycle. To keep your balance, you must keep moving." - 
Albert Einstein



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Re: [Histonet] Question - Bone marrow core biopy specimens

[Victoria Baker via Histonet]

Hi Richard!

That is the duplicate of what we do in our lab.  We were putting both the
core and the clot on the same slide for in house testing, but den outs we
sometimes have to split the specimen and only on a slide.

I have a question for you about ISH Kappa/Lambda.  What decal protocol do
you use and do you see edge non specific detection/substrate on your
slides?  We use the Roche Iview blue.

I hope what I said helps.

Vikki

On Mon, May 3, 2021 at 4:12 PM Cartun, Richard via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Hello everyone:
>
> I'm wonder what your protocol is for cutting formalin-fixed,
> paraffin-embedded bone marrow core biopsy specimens?  Do you cut a limited
> number of slides upfront and then go back to the block if the pathologist
> requests histochemical stains or immunohistochemical tests?  We are
> implementing a protocol today where we will cut "15" slides upfront; 3 H
> at different levels and 12 unstains for potential histochemical stains
> and/or IHC tests.  Are we crazy?
>
> Thank you, and I hope everyone is doing well.
>
> Richard
>
> Richard W. Cartun, MS, PhD
> Director, Histology & The Martin M. Berman, MD Immunopathology/Morphologic
> Proteomics Laboratory
> Director, Biospecimen Collection Programs
> Assistant Director, Anatomic Pathology
> Hartford Hospital
> 80 Seymour Street
> Hartford, CT  06102
> (860) 972-1596 (Office)
> (860) 545-2204 (Fax)
> richard.car...@hhchealth.org
>
>
> This e-mail message, including any attachments, is for the sole use of the
> intended recipient(s) and may contain confidential and privileged
> information. Any unauthorized review, use, disclosure, or distribution is
> prohibited. If you are not the intended recipient, or an employee or agent
> responsible for delivering the message to the intended recipient, please
> contact the sender by reply e-mail and destroy all copies of the original
> message, including any attachments.
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[Histonet] Question - Bone marrow core biopy specimens

[Cartun, Richard via Histonet]

Hello everyone:

I'm wonder what your protocol is for cutting formalin-fixed, paraffin-embedded 
bone marrow core biopsy specimens?  Do you cut a limited number of slides 
upfront and then go back to the block if the pathologist requests histochemical 
stains or immunohistochemical tests?  We are implementing a protocol today 
where we will cut "15" slides upfront; 3 H at different levels and 12 
unstains for potential histochemical stains and/or IHC tests.  Are we crazy?

Thank you, and I hope everyone is doing well.

Richard

Richard W. Cartun, MS, PhD
Director, Histology & The Martin M. Berman, MD Immunopathology/Morphologic 
Proteomics Laboratory
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596 (Office)
(860) 545-2204 (Fax)
richard.car...@hhchealth.org


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Re: [Histonet] Question about accessioning outside consult cases

[Lindrud, Scott via Histonet]

Hi Martha,

We have been using Beaker for over 2 years now.  We created another accession 
that identifies cases that are being sent to us as a consult/referral (both 
Cyto and Histo).   It works pretty well for us.

Scott


Scott A. Lindrud, MLS(ASCP)CT  | Histopathology Technical 
Specalist/Cytotechnologist

P: 320-231-4520
F: 320-231-4503
scott.lind...@carrishealth.com

Carris-Health
Rice Memorial Hospital
301 Becker Ave SW
Willmar, MN 56201






-Original Message-
From: Martha Ward-Pathology 
Sent: Thursday, December 10, 2020 12:45 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Question about accessioning outside consult cases

We are in the process of switching to AP Beaker and in the midst of the build.  
  My question involves how others are handling outside consult surgical and/or 
cytology cases.   Do you assign them an unique number that identifies them as a 
consult as opposed to an inside case - SO20- verses S20-? We 
currently assign them as just another surgical case but I know some places do 
have outside consult designations.

What is everyone doing?

Thanks in advance for your input.

Martha Ward, MT ASCP (QIHC)
Manager, Molecular Diagnostics Lab
Wake Forest Baptist Medical Center
Winston-Salem, NC 27157

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Re: [Histonet] Question about accessioning outside consult cases

[Morken, Timothy via Histonet]

Martha, we give it the same number sequence as any other surgical or cytology 
case, but we can identify our cases by "Specimen Class" so consults get a 
specimen class according to the type of consult - cyto,  cytogyn, surgical, 
etc.  The specimen class is printed on the labels for all materials.

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

-Original Message-
From: Martha Ward-Pathology via Histonet  
Sent: Thursday, December 10, 2020 10:45 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Question about accessioning outside consult cases

We are in the process of switching to AP Beaker and in the midst of the build.  
  My question involves how others are handling outside consult surgical and/or 
cytology cases.   Do you assign them an unique number that identifies them as a 
consult as opposed to an inside case - SO20- verses S20-? We 
currently assign them as just another surgical case but I know some places do 
have outside consult designations.

What is everyone doing?

Thanks in advance for your input.

Martha Ward, MT ASCP (QIHC)
Manager, Molecular Diagnostics Lab
Wake Forest Baptist Medical Center
Winston-Salem, NC 27157
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Re: [Histonet] Question about accessioning outside consult cases

[Lester Raff via Histonet]

We identify our outside surgical consults as OC, for example OC20-00010
Les Raff




On Thu, Dec 10, 2020 at 12:45 PM Martha Ward-Pathology via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> We are in the process of switching to AP Beaker and in the midst of the
> build.My question involves how others are handling outside consult
> surgical and/or cytology cases.   Do you assign them an unique number that
> identifies them as a consult as opposed to an inside case - SO20-
> verses S20-? We currently assign them as just another surgical case
> but I know some places do have outside consult designations.
>
> What is everyone doing?
>
> Thanks in advance for your input.
>
> Martha Ward, MT ASCP (QIHC)
> Manager, Molecular Diagnostics Lab
> Wake Forest Baptist Medical Center
> Winston-Salem, NC 27157
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[Histonet] Question about accessioning outside consult cases

[Martha Ward-Pathology via Histonet]

We are in the process of switching to AP Beaker and in the midst of the build.  
  My question involves how others are handling outside consult surgical and/or 
cytology cases.   Do you assign them an unique number that identifies them as a 
consult as opposed to an inside case - SO20- verses S20-? We 
currently assign them as just another surgical case but I know some places do 
have outside consult designations.

What is everyone doing?

Thanks in advance for your input.

Martha Ward, MT ASCP (QIHC)
Manager, Molecular Diagnostics Lab
Wake Forest Baptist Medical Center
Winston-Salem, NC 27157
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[Histonet] Question for Histonet

[Sharon Wheeler via Histonet]

I was wondering if other facilities have the same issue as us?





We have excess breakage (brittleness) of the solution reservoirs (680 mL) for 
our PRISMA stainer. We clean the dirty reservoirs in an automatic dishwasher 
with a 18 minute dry cycle at 90 degrees.

How do other places clean their baskets? Has anybody else had similar 
experience???


Thanks,
Sharon

Senior Technologist Surgical & Ancillary Pathology
Pathology and Laboratory Medicine
London Health Sciences Centre and St. Joseph's Health Care London
University Hospital, Rm A3-236
TEL:  519-685-8500, ext. 34711
E-mail: sharon.whee...@lhsc.on.ca


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Re: [Histonet] question on processing worms (1...@comcast.net)

[Hobbs, Carl via Histonet]

Hi
I have done several yrs of work on Eisenia fetida worms with a PhD student.
I decided to do a std Pwax processing schedule.
The student dissected the appropriate area then placed in 10% Formalin in PBS 
pH7.4 for 24hrs on a rocker ( gentle agitation)
I then processed to Pwax using a std IMS/Xylene/wax on a Leica carousel ( 1hr 
in each station)
Worked fine.
Could do tinctorial and IHC no problem.
Sure, many primary Abs didn't work...
We also did TUNEL on them Pwax sectionsworked a treat.
I may have TUNEL images in the Histonet Image archive.

Good luck!
NB: Sure, maybe freezing is a better way forwards. Depends. 
All I know is that the PhD student passed with no problems and went on to a so 
far succesful career in Science Industry

Best wishes

Carl

Carl Hobbs FIBMS
Histology and Imaging Manager
Wolfson CARD
Guys Campus, London Bridge 
Kings College London
London
SE1 1UL
 


020 7848 6813
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[Histonet] question on processing worms

[LEROY H BROWN via Histonet]

I have crassicauda specimens ( kidney worms - relatively large worms 1-2 mm
width) And would like to know how you fix and process these for paraffin
sections of H slides.Do you do anything different for this?

Thanks

LeRoy Brown HT(ASCP) HTL 

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Re: [Histonet] Question concerning H. pylori staining

[jkiernan--- via Histonet]

In the method of Leung et al. (1996 J. Histotechnol. 19:131), the periodic acid 
oxidizes the neutral gastric mucus to generate aldehyde groups, as in the first 
part of the PAS method. Treatment with an acidified bisulphite solution then 
converts the aldehyde groups to aldehyde bifulphite groups, which are strong 
anions. Alcian yellow (a cationic azo dye) binds strongly to these anionic 
sites (but not to nucleic acids) and may then change into a permanently 
insoluble pigment. (The chemistry is similar to that of staining with alcian 
blue 8G; this isn't the place to discuss all that!)

The next step (mildly alkaline aqueous toluidine blue) imparts blue to nearly 
everything not already coloured yellow; it cannot displace alcian yellow from 
the already stained neutral gastric mucus, making for strong contrast. The 
bacteria stain blue by virtue of their anionic components (nucleic acids and 
materials in their cell walls) so do nuclei of cells. Toluidine blue, like most 
dyes with small molecules, stains quickly and is also easy to remove. 
Alcohol-water mixtures remove such dyes faster than either water or 100% 
alcohol. The original Leung method recommended removing most of the water by 
careful blotting, and then taking the slide(s) directly to the first of 3 
changes of 100% alcohol.

Real alcian yellow (CI 12840, Ingrain yellow 1) has not been manufactured for 
many years. Other methods for showing blue bacilli in yellow mucus are 
available, but the chemistry for getting the yellow is not the same.

John Kiernan
Anatomy, UWO, London, Canada
https://www.schulich.uwo.ca/anatomy/people/bios/emeriti/kiernan_john.html
= = =

From: Gudrun Lang via Histonet 
Sent: 12 April 2020 03:38
To: histonet@lists.utsouthwestern.edu 
Subject: Re: [Histonet] Question concerning H. pylori staining

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

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

Gudrun Lang



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

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

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

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

Bob Richmond
Samurai Pathologist
Maryville TN
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Re: [Histonet] Question concerning H. pylori staining

[Gudrun Lang via Histonet]

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

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

Gudrun Lang



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

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

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

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

Bob Richmond
Samurai Pathologist
Maryville TN
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Re: [Histonet] Question concerning H. pylori staining

[Bob Richmond via Histonet]

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

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

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

Bob Richmond
Samurai Pathologist
Maryville TN
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[Histonet] Question concerning H-Pylori Staining

[Ken M via Histonet]

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

Michelle
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Re: [Histonet] question about Digital Pathology Certificate

[Ranna via Histonet]

Thank you Victoria 
Regards 
Ranna

Sent from my iPhone

> On Apr 8, 2020, at 4:39 AM, Victoria Baker  wrote:
> 
> 
> Ranna,
> 
> I took the course and it was well worth the cost for me.  As to how it is 
> assisting my career right now it isn't as my facility stopped using imaging 
> about a year and a half ago.  With the pandemic they may be rethinking this, 
> but it won't be immediate.
> This type of skill will be more important, I believe, with the advancement of 
> telemedicine and what we should be learning from the COVID-19 shut down or 
> limited resources of certain services.  Plus with so many facilities merging 
> to survive in the current environment this is going to be necessary.
> 
> What I would like to see is more follow up courses and more attention drawn 
> to this program.
> 
> Vikki
> 
> 
> 
>> On Mon, Apr 6, 2020, 7:48 PM Ranna Mehta via Histonet 
>>  wrote:
>> Hi All,
>> I need inputs from my histo colleagues about Digital Pathology
>> certification from NSH. If anyone has done this program, how is it helping
>> in their career?
>> what are pros and cons? Do they include MatLab  and R programming in
>> syllabus? I know it is very resourceful in research lab but what about
>> Clinical labs?
>> Right now i am working on multiplex staining using Vectra inform and Halo.
>> I am self learner; I don't have any help except company's technical
>> support. I am wondering- attending this program will help me understanding
>> algorithm and analysis? Thanks in advanced. Stay safe.
>> sorry for lots of questions.
>> Regards
>> Ranna Mehta
>> Associate Scientist
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Re: [Histonet] question about Digital Pathology Certificate

[Victoria Baker via Histonet]

Ranna,

I took the course and it was well worth the cost for me.  As to how it is
assisting my career right now it isn't as my facility stopped using imaging
about a year and a half ago.  With the pandemic they may be rethinking
this, but it won't be immediate.
This type of skill will be more important, I believe, with the advancement
of telemedicine and what we should be learning from the COVID-19 shut down
or limited resources of certain services.  Plus with so many facilities
merging to survive in the current environment this is going to be necessary.

What I would like to see is more follow up courses and more attention drawn
to this program.

Vikki



On Mon, Apr 6, 2020, 7:48 PM Ranna Mehta via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Hi All,
> I need inputs from my histo colleagues about Digital Pathology
> certification from NSH. If anyone has done this program, how is it helping
> in their career?
> what are pros and cons? Do they include MatLab  and R programming in
> syllabus? I know it is very resourceful in research lab but what about
> Clinical labs?
> Right now i am working on multiplex staining using Vectra inform and Halo.
> I am self learner; I don't have any help except company's technical
> support. I am wondering- attending this program will help me understanding
> algorithm and analysis? Thanks in advanced. Stay safe.
> sorry for lots of questions.
> Regards
> Ranna Mehta
> Associate Scientist
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
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[Histonet] question about Digital Pathology Certificate

[Ranna Mehta via Histonet]

Hi All,
I need inputs from my histo colleagues about Digital Pathology
certification from NSH. If anyone has done this program, how is it helping
in their career?
what are pros and cons? Do they include MatLab  and R programming in
syllabus? I know it is very resourceful in research lab but what about
Clinical labs?
Right now i am working on multiplex staining using Vectra inform and Halo.
I am self learner; I don't have any help except company's technical
support. I am wondering- attending this program will help me understanding
algorithm and analysis? Thanks in advanced. Stay safe.
sorry for lots of questions.
Regards
Ranna Mehta
Associate Scientist
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Re: [Histonet] Question about gelatin embedding

[John Kiernan via Histonet]

Gelatin embedding is easy. You infiltrate the specimen and then fix it again in 
formaldehyde to cross-link the gelatin molecules and make the whole mass 
isoluble in water. You can than cut frozen sections of any kind: cryostat, or 
with an old-fashioned freezing microtome collecting thawed sections from the 
knife with a brush.  The formaldehyde-fixed gelatin holds everything together.  
With a Nissl stain it remains inconspicuous. If you do an H the gelatin will 
stain red.
John Kiernan
= = =

From: Alonso Martínez Canabal via Histonet 
Sent: 17 January 2020 16:29
To: histonet@lists.utsouthwestern.edu 
Subject: [Histonet] Question about gelatin embedding

Hello,
  I am here again. I am wondering if someone has good experience
embedding in gelatin-albumin for cryostat or vibratome sectioning.
Specifically we use brain tissue and is common in free floating techniques
non-attached parts of the same section float around and later that
generates all sorts of problems.
   Thank you very much.

--
Dr. Alonso Martínez Canabal PhD
Profesor Asociado "C"
Departamento de Biología Celular, Facultad de Ciencias, UNAM
Investigador Nacional "I"
56224833
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[Histonet] Question about gelatin embedding

[Alonso Martínez Canabal via Histonet]

Hello,
  I am here again. I am wondering if someone has good experience
embedding in gelatin-albumin for cryostat or vibratome sectioning.
Specifically we use brain tissue and is common in free floating techniques
non-attached parts of the same section float around and later that
generates all sorts of problems.
   Thank you very much.

-- 
Dr. Alonso Martínez Canabal PhD
Profesor Asociado "C"
Departamento de Biología Celular, Facultad de Ciencias, UNAM
Investigador Nacional "I"
56224833
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[Histonet] Question for Leica Bond IHC Users

[Cartun, Richard via Histonet]

We are preparing to install a Leica "Bond Stain Interface" which will allow us 
to move away from having to print slide labels manually for our IHC slides.  I 
am not an expert on this so I am looking for help.  We have "5" Bond IHC 
instruments that will be hooked-up to the controller; however, we have been 
told that we cannot connect our 6th instrument unless we get a second 
controller (and that's not going to happen).  Has anyone else been in a similar 
predicament?  And, if so, how have you resolved it?  We need to use all "6" 
instruments.  Thank you.

Richard

Richard W. Cartun, MS, PhD
Director, Histology & The Martin M. Berman, MD Immunopathology/Morphologic 
Proteomics Laboratory
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596 (Office)
(860) 545-2204 (Fax)
richard.car...@hhchealth.org


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Re: [Histonet] Question - EM

[Greg Dobbin via Histonet]

Hi Dr. Cartun,
It has been many years since I worked in EM but I my recollection is that
tissues could remain in 2% Glut indefinitely without detriment (for EM
purposes). However, Osmium tetroxide had to have a limited exposure.
Greg

-- 
*Greg Dobbin*
1205 Pleasant Grove Rd
RR#2 York,
PE  C0A 1P0


*Everything in moderation...even moderation itself**!*
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[Histonet] Question - EM

[Cartun, Richard via Histonet]

How long can tissue remain in glutaraldehyde before EM testing is performed?  
Thank you.

Richard

Richard W. Cartun, MS, PhD
Director, Histology & The Martin M. Berman, MD Immunopathology & Morphologic 
Proteomics Laboratory
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596
(860) 545-2204 Fax


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[Histonet] Question regarding frozen control tissue

[Haley Huggins via Histonet]

I am trying to find a way to obtain small bowel/intestine as a fresh/frozen
sample to be able to use it for a control with my staining for PGP9.5 free
floating IHC stain. Does anyone know where a good place would be to obtain
the needed control tissue?

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

[Cartun, Richard via Histonet]

We are thinking of changing the way we report consult testing (IHC, Flow 
Cytometry, and Molecular) for our system hospitals.  Instead of accessioning 
the consult case here, the requesting hospital would create a 
"Procedure/Addendum" in Sunquest CoPath for their specimen and then send us the 
unstained slides or paraffin block for testing.  Once the testing is completed, 
the pathologist here (assigned to the case) would go into the other hospital's  
report, populate the "Procedure/Addendum" with the testing performed and the 
interpretation, and then sign it out.  We would not have a consult accession 
number (or report) here at our hospital for this specimen.  Does this make 
sense?  Is anyone else in a multiple hospital system already doing this?  I 
welcome all comments.  Thank you.

Richard

Richard W. Cartun, MS, PhD
Director, Histology & The Martin M. Berman, MD Immunopathology & Morphologic 
Proteomics Laboratory
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596
(860) 545-2204 Fax


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Re: [Histonet] Question on Sanderson Rapid Bone Stain

[Jessica Riggleman via Histonet]

RBS is the same as Methylene Blue, I believe.



_

Jessica Riggleman | Research Associate

Globus Medical, Inc.
Valley Forge Business Center
2560 General Armistead Avenue | Audubon, PA 19403
Ph: (610) 930-1800 ext. 2583 | Fax:

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-Original Message-
From: Alicia Marie Ortega [mailto:alicia.ort...@colorado.edu]
Sent: Thursday, September 22, 2016 4:50 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Question on Sanderson Rapid Bone Stain

Hello everyone,


I am attempting to stain 30-40 micron thick undecalcified bone sections 
embedded in poly(methyl methacrylate) with Sanderson Rapid Bone Stain (RBS) 
with a Van Geison counter stain.


My first attempt at this stain resulted in a very faint stain from the RBS (I 
could see very faint blue/green staining of osteoid/soft tissues) and a very 
intense dark pink stain from the counterstain.  I was wondering if anyone knows 
of a way to intensify the staining of the RBS?  I heated the stain to 55-60 
degrees Celsius as recommended by the manufacturer prior to staining (with a 10 
minute stain duration).


Thank you in advance for your time and help.


Sincerely,

Alicia Ortega

Postdoctoral Research Associate

University of Colorado, Boulder

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[Histonet] Question on Sanderson Rapid Bone Stain

[Alicia Marie Ortega via Histonet]

Hello everyone,


I am attempting to stain 30-40 micron thick undecalcified bone sections 
embedded in poly(methyl methacrylate) with Sanderson Rapid Bone Stain (RBS) 
with a Van Geison counter stain.


My first attempt at this stain resulted in a very faint stain from the RBS (I 
could see very faint blue/green staining of osteoid/soft tissues) and a very 
intense dark pink stain from the counterstain.  I was wondering if anyone knows 
of a way to intensify the staining of the RBS?  I heated the stain to 55-60 
degrees Celsius as recommended by the manufacturer prior to staining (with a 10 
minute stain duration).


Thank you in advance for your time and help.


Sincerely,

Alicia Ortega

Postdoctoral Research Associate

University of Colorado, Boulder
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Re: [Histonet] Question about prelim report for autopsy with neuropathology

[Megan Dishop via Histonet]

Hi Tim, 
In the hospitals I have worked, we used the PAD simply to communicate gross 
pathologic diagnoses, so we would issue a gross description of the brain (like 
the brain weight compared to normal, edema, atrophy, herniation, hemorrhage, 
etc), "pending microscopic examination", and then follow with the FAD after 
microscopic exam is done.  It might just be a few lines, but that's OK.  It 
gives the clinical team a quick update of what was observed and what is pending.

Megan
 


Megan K. Dishop MD
Medical Director, Pediatric Anatomic Pathology
Children's Hospitals and Clinics of Minnesota Laboratories
2525 Chicago Ave S. MS32-B600, Minneapolis, MN 55404 USA
Phone: 612-813-6521Fax: 612-813-7721  Email: megan.dis...@childrensmn.org
Adjoint Professor of Pediatrics, University of Colorado School of Medicine




>>> "Morken, Timothy via Histonet"  
>>> 7/29/2016 4:12 PM >>>
How do you handle counting days to preliminary autopsy dx (PAD) when doing 
cases in which the brain autopsy results are the main component of disease? If 
we accession at the time of autopsy and then let the brain fix for several days 
before cutting in, then the PAD is many days past the 3-day reporting 
requirement.

On standard "body" cases we report PAD the same day and final within two weeks. 
With neuropathology cases it can be up to 10 days for a PAD.

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

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[Histonet] Question about prelim report for autopsy with neuropathology

[Morken, Timothy via Histonet]

How do you handle counting days to preliminary autopsy dx (PAD) when doing 
cases in which the brain autopsy results are the main component of disease? If 
we accession at the time of autopsy and then let the brain fix for several days 
before cutting in, then the PAD is many days past the 3-day reporting 
requirement.

On standard "body" cases we report PAD the same day and final within two weeks. 
With neuropathology cases it can be up to 10 days for a PAD.

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

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[Histonet] Question - CoPath

[Cartun, Richard via Histonet]

We will be switching from Cerner CoPath to Sunrise CoPath in the near future 
(part of our Epic conversion).  Is there anyone using Sunrise CoPath that uses 
a prostate diagram in their report to demonstrate the absence or presence of 
cancer in the different quadrants?  Thank you.

Richard

Richard W. Cartun, MS, PhD
Director, Histology & The Martin M. Berman, MD Immunopathology & Morphologic 
Proteomics Laboratory
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596
(860) 545-2204 Fax


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[Histonet] Question About A Sponge Type

[Marcum, Pamela A via Histonet]

When I was at NSH in Austin, I talked to a vendor with a very different type of 
Histology.  It was very soft and far less porous than the normal blue sponges.  
Does anyone remember this or can you tell me where to get them?  I believe the 
company was from the UK with offices or distributors in the US.  I need to 
either buy or get some for a demo here with our pathologists.  We cannot use 
the blue sponges for many of our cases as they just too rough and the surfaces 
are too rough.

Thank You,

Pam Marcum
UAMS

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Re: [Histonet] Question re: accessory piece for tissue flotation bath

[Manfre, Philip via Histonet]

With regards to the HistoOrientator, it does removes wrinkles from the sections 
fairly effectively.  That being said, I would not use it on any sections 
intended for immunohistochemistry.  The hot plate on this device gets very hot 
and would no doubt damage or affect the antigenicity of such tissues.  Also, 
some practice is required to effectively use it.  If you do not drain the 
slides somewhat first, the tissues will quickly move all over the slide as they 
heat.  They will move quite a bit, sometimes butting up against each other.  If 
you drain them too long, then the tissues start to dry onto the slide, wrinkles 
and all.  So, as I said, spend some time practicing with this device with 
practice tissue until you get the hang of it.  It is a good tool that is tricky 
to use at first.

Good luck!
Phil.

Philip Manfre, B.A., HT (ASCP)
Associate Principal Scientist
Merck Research Laboratories
WP45-251
PO Box 4
West Point, PA 19486

215-652-9750
215-993-0383 (fax)
philip_man...@merck.com



-Original Message-
From: Sanders, Jeanine (CDC/OID/NCEZID) via Histonet 
[mailto:histonet@lists.utsouthwestern.edu] 
Sent: Wednesday, February 24, 2016 8:36 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Question re: accessory piece for tissue flotation bath

Morning!

Can anyone of you share the functionality of:

Flotation Work Station w/ 8" x 8" x 2 1/4" (Deep Dish), HistoOrientator and 
Dryer

Curious if the HistoOrientator actually removes wrinkles as the description 
states.

Thanks!

Jeanine H. Sanders, BS, HT (ASCP), QIHC
Centers for Disease Control and Prevention
1600 Clifton Rd., NE MS/G-32
Atlanta, GA 30329
j...@cdc.gov<mailto:j...@cdc.gov>
404-639-3590

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[Histonet] Question re: accessory piece for tissue flotation bath

[Sanders, Jeanine (CDC/OID/NCEZID) via Histonet]

Morning!

Can anyone of you share the functionality of:

Flotation Work Station w/ 8" x 8" x 2 1/4" (Deep Dish), HistoOrientator and 
Dryer

Curious if the HistoOrientator actually removes wrinkles as the description 
states.

Thanks!

Jeanine H. Sanders, BS, HT (ASCP), QIHC
Centers for Disease Control and Prevention
1600 Clifton Rd., NE MS/G-32
Atlanta, GA 30329
j...@cdc.gov
404-639-3590

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Re: [Histonet] question - Allergy to histological solvents?

[Rene J Buesa via Histonet]

You can dewax absolutely safely using a 2% dishawasher soap solution at 90ºC 
(twice) as washing in water.You can "dehydrate" stained stains by placing the 
slides in an oven at 60ºC, also absolutely safely for the stained section.Under 
separate cover I am sending  articles on this subject.René 

On Saturday, February 20, 2016 3:05 PM, daniel blackburn via Histonet 
 wrote:
 

 Hello

After decades of using standard organic solvents for paraffin- section 
histology, I find that I've become highly allergic to the fumes.  These include 
commercial preps like Citrosolv and Hemo-De, as well as xylene and toluene, 
which cause (in me) dizziness and a pounding headache.  This problem has made 
staining and coverslipping very difficult, even with  hood.  Can anyone 
recommend alternative solutions for (a) dewaxing prior to staining and (b) 
coverslipping?  thanks very much!

Daniel Blackburn
Biology, Trinity College (Hartford CT)

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[Histonet] question - Allergy to histological solvents?

[daniel blackburn via Histonet]

Hello

After decades of using standard organic solvents for paraffin- section 
histology, I find that I've become highly allergic to the fumes.  These include 
commercial preps like Citrosolv and Hemo-De, as well as xylene and toluene, 
which cause (in me) dizziness and a pounding headache.  This problem has made 
staining and coverslipping very difficult, even with  hood.   Can anyone 
recommend alternative solutions for (a) dewaxing prior to staining and (b) 
coverslipping?  thanks very much!

Daniel Blackburn
Biology, Trinity College (Hartford CT)

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[Histonet] Question for General Data users

[Amanda Reichard via Histonet]

Good Morning!

Does anyone else use General Data for their barcode labeling system?  If you 
do, is your operating system the HTS system or Lab Cycle?

Thanks in advance!
Amanda Reichard, HTL (ASCP)cm
Histology Supervisor
Laboratory
Licking Memorial Health Systems
1320 W. Main St.
Newark, OH 43055
(740) 348-4163


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Re: [Histonet] Question

[Goins, Tresa]

All that matters here is the final concentration of the reagent - it doesn't 
matter what stock you start with if you calculate the dilution.
Adding 1.25 ml to 1000 ml is diluted by a factor of .00125 so 10N x .00125 = 
.0125 N final concentration.
If using a 1 N stock solution, just add 10X the volume or 12.5 ml 1 N stock to 
987.5 ml water.

Hope this helps,

Tresa

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bernice 
Frederick
Sent: Thursday, April 30, 2015 1:35 PM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Question

All,
I have a procedure here that call for and I quote 1.25 ml NaOH 10N in 1L of 
water. I know how to get 1 N, but how do I get 10. Having rarely hd the 
opportunity to make many Normal solutions ,my brain is not computing. Is it an 
error?
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.edumailto:b-freder...@northwestern.edu

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Re: [Histonet] Question

[Geoff]

Huh?
Take a solution more dilute than you want it and dilute it more?

Geoff

On 5/1/2015 10:41 AM, Goins, Tresa wrote:

All that matters here is the final concentration of the reagent - it doesn't 
matter what stock you start with if you calculate the dilution.
Adding 1.25 ml to 1000 ml is diluted by a factor of .00125 so 10N x .00125 = 
.0125 N final concentration.
If using a 1 N stock solution, just add 10X the volume or 12.5 ml 1 N stock to 
987.5 ml water.

Hope this helps,

Tresa

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bernice 
Frederick
Sent: Thursday, April 30, 2015 1:35 PM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Question

All,
I have a procedure here that call for and I quote 1.25 ml NaOH 10N in 1L of 
water. I know how to get 1 N, but how do I get 10. Having rarely hd the opportunity 
to make many Normal solutions ,my brain is not computing. Is it an error?
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.edumailto:b-freder...@northwestern.edu

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**
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732) 235-4583; fax: -4029
mcaul...@rwjms.rutgers.edu
**



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Re: [Histonet] Question

[Goins, Tresa]

The final concentration of 1.25 ml 10N NaOH into 1000 ml water is the same as:
12.5 ml 1N NaOH into 987.5 ml water.

-Original Message-
From: Geoff [mailto:mcaul...@rwjms.rutgers.edu] 
Sent: Friday, May 01, 2015 9:16 AM
To: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Question

Huh?
Take a solution more dilute than you want it and dilute it more?

Geoff

On 5/1/2015 10:41 AM, Goins, Tresa wrote:
 All that matters here is the final concentration of the reagent - it doesn't 
 matter what stock you start with if you calculate the dilution.
 Adding 1.25 ml to 1000 ml is diluted by a factor of .00125 so 10N x .00125 = 
 .0125 N final concentration.
 If using a 1 N stock solution, just add 10X the volume or 12.5 ml 1 N stock 
 to 987.5 ml water.

 Hope this helps,

 Tresa

 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu 
 [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bernice 
 Frederick
 Sent: Thursday, April 30, 2015 1:35 PM
 To: Histonet@lists.utsouthwestern.edu
 Subject: [Histonet] Question

 All,
 I have a procedure here that call for and I quote 1.25 ml NaOH 10N in 1L of 
 water. I know how to get 1 N, but how do I get 10. Having rarely hd the 
 opportunity to make many Normal solutions ,my brain is not computing. Is it 
 an error?
 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.edumailto:b-freder...@northwestern.edu

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--
**
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732) 235-4583; fax: -4029
mcaul...@rwjms.rutgers.edu
**



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[Histonet] Question

[Bernice Frederick]

All,
I have a procedure here that call for and I quote 1.25 ml NaOH 10N in 1L of 
water. I know how to get 1 N, but how do I get 10. Having rarely hd the 
opportunity to make many Normal solutions ,my brain is not computing. Is it an 
error?
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.edumailto:b-freder...@northwestern.edu

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[Histonet] Question on IHC billing

[Vickroy, James]

Let me see if I have this straight:If a pathologist orders an Hpylori stain 
on 2 blocks from the same specimen C1 and C2 we can only bill one 88342.

If this correct.Obviously if he ordered addition different IHC stains we 
could change additional 88341's.

Jim Vickroy
Histology Manager
Springfield Clinic, Main Campus, East Building
1025 South 6th Street
Springfield, Illinois  62703
Office:  217-528-7541, Ext. 15121
Email:  jvick...@springfieldclinic.commailto:jvick...@springfieldclinic.com



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[Histonet] Question about Formic acid decal and TRAP stain

[Debra Siena]

Hi Histonetters,

I have a question to ask if you don't mind.  Can TRAP Histochemical staining be 
performed after decalcifying with formic acid? Any tricks of the trade, etc?  
If anyone has any experience or references that they could point me to, I would 
greatly appreciate it.  Thanks in advance for your help.




Debbie Siena
dsi...@statlab.com mailto:bbro...@statlab.com%7C | 
www.statlab.comhttp://www.statlab.com/

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Re: [Histonet] Question about Formic acid decal and TRAP stain

[koellingr]

Hi Debra, 
My experience differs from Elizabeth and others.  Indeed have made thousands of 
mouse bone preparations with formic acid decaled sections.  Here is an article 
that didn't copy paste so well: 
  


RANK is the intrinsic hematopoietic cell surface 

receptor that controls osteoclastogenesis and 

regulation of bone mass and calcium metabolism 

Ji Li*, Ildiko Sarosi 

† , Xiao-Qiang Yan † , Sean Morony † , Casey Capparelli † , Hong-Lin Tan † 

, Susan McCabe*, Robin Elliott*, 

Sheila Scully 

 Gwyneth Van † , Stephen Kaufman † , Shao-Chieh Juan † , Yu Sun † , John 
Tarpley † , Laura Martin † 

, Kathleen Christensen 

 James McCabe † , Paul Kostenuik † , Hailing Hsu*, Frederick Fletcher † , Colin 
R. Dunstan † 

, David L. Lacey 

, and William J. Boyle* 
Departments of *Cell Biology and 

Pathology, Amgen Inc., One Amgen Center Drive, Thousand Oaks, CA 91320 

Edited by David V. Goeddel, Tularik, Inc., South San Francisco, CA, and 
approved December 20, 1999 (received for review September 30, 1999) 

  

PNAS 

u February 15, 2000 u vol. 97 u no. 4 u 

1571 

  

Is in PNAS and while the methods are not here in this article, look in the 
reference section to materials and methods used and see beautiful pictures of 
TRAP from formic acid decaled bones.  I did them for years after that.  
Complete fixation and gentle formic acid (or immunocal) and indeed this can be 
done. 

  

Ray (retired in Lake Forest Park, WA, golf and science education outreach for 
K-12 busy) 










  
- Original Message -

From: Debra Siena dsi...@statlab.com 
To: histonet@lists.utsouthwestern.edu 
Sent: Thursday, April 2, 2015 9:00:25 AM 
Subject: [Histonet] Question about Formic acid decal and TRAP stain 

Hi Histonetters, 

I have a question to ask if you don't mind.  Can TRAP Histochemical staining be 
performed after decalcifying with formic acid? Any tricks of the trade, etc?  
If anyone has any experience or references that they could point me to, I would 
greatly appreciate it.  Thanks in advance for your help. 




Debbie Siena 
dsi...@statlab.com mailto:bbro...@statlab.com%7C | 
www.statlab.comhttp://www.statlab.com/ 

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[Histonet] Question about POC testing

[Pratt, Caroline]

Does anyone know if a Tzanck Smear falls under Virology or Cytology for POC 
testing in a Dermatology clinic.  I have heard conflicting theories.  Thanks!


Caroline M. Pratt, MBA
Practice Administrator Dermpath
3020 Market Street, Ste 201
Philadelphia, PA  19104
Phone 215-349-8178
Cell 610-800-1381
Fax 215-662-6150

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RE: [Histonet] Question about POC testing

[suetp918]

Cyto


Sent from my Verizon Wireless 4G LTE smartphone


 Original message 
From: Pratt, Caroline caroline.pr...@uphs.upenn.edu 
Date:02/19/2015  4:59 PM  (GMT-05:00) 
To: 'histonet' histonet@lists.utsouthwestern.edu 
Subject: [Histonet] Question about POC testing 

Does anyone know if a Tzanck Smear falls under Virology or Cytology for POC 
testing in a Dermatology clinic.  I have heard conflicting theories.  Thanks!


Caroline M. Pratt, MBA
Practice Administrator Dermpath
3020 Market Street, Ste 201
Philadelphia, PA  19104
Phone 215-349-8178
Cell 610-800-1381
Fax 215-662-6150

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[Histonet] Question regarding dehydration

[Julio Benavides]

Hi there,

we are having problems trying to cut some embedded samples (they crumble 
in the bath and the few cuts we manage to get into HE are crap). These 
are formalin fixed samples (bovine foetal and placenta samples) which 
went straight from formlin into 100º ethanol for the dehydration before 
clearing. I was wondering if such drastic dehydration step (no 60º, 70º 
or 90º ethanol before the 100º) could have damage the tissue. Has anyone 
have a similar issue before? do you think the samples are ruined for 
histology?


Thanks a lot for your help

Regards

Julio
Instituto de Ganaderia de Montaña
Spain

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RE: [Histonet] Question regarding dehydration

[Julio Benavides]

Hi Ryan,

thanks a lot for your thoughts. These blocks were processed elsewhere 
and sent to us for the cutting and staining. Tissues were dehydrated in 
five consecutive baths of ethanol 100%, 5 hours each (manual 
processing). Then, they went to xilene (three baths, 4 hours each) and 
paraffin (two batch, 1h30´ each). Truth is that I have never seen such 
processing before. In our lab, with automatic processing, we begin with 
60% ethanol and go to 70, then 95 and then 100 before ethanol/xylene.


I was wondering if anybody has used the 100% ethanol processing before 
and which was the influence over tissues.


Thanks

Julio


 Forwarded Message 
Subject:RE: [EXTERNAL] [Histonet] Question regarding dehydration
Date:   Fri, 13 Feb 2015 09:06:47 -0500
From:   Roy, Ryan ryan@va.gov
To: 'Julio Benavides' j.benavi...@eae.csic.es



If the tissue is fatty it will blow apart on the water bath. If the H and E 
staining is poor its probably the processing.

What is the exact procedure for your process including time that the tissue was 
in each reagent? I would include this in a message to all of histonet as well 
as some may more knowledgeable than myself.

Also, I would recommend using a 95% solution prior to the 100% alcohol (s) if 
not a 70-80% and 95%.


Ryan Roy HTL (ASCP)
Manchester Veterans Affairs Medical Center
Manchester New Hampshire

Disclosure: The content of this email does not represent the views or opinons 
of the VA





-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Julio Benavides
Sent: Friday, February 13, 2015 4:26 AM
To: histonet@lists.utsouthwestern.edu
Subject: [EXTERNAL] [Histonet] Question regarding dehydration

Hi there,

we are having problems trying to cut some embedded samples (they crumble in the 
bath and the few cuts we manage to get into HE are crap). These are formalin 
fixed samples (bovine foetal and placenta samples) which went straight from 
formlin into 100º ethanol for the dehydration before clearing. I was wondering 
if such drastic dehydration step (no 60º, 70º or 90º ethanol before the 100º) 
could have damage the tissue. Has anyone have a similar issue before? do you 
think the samples are ruined for histology?

Thanks a lot for your help

Regards

Julio
Instituto de Ganaderia de Montaña
Spain

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Re: [EXTERNAL] RE: [Histonet] Question regarding dehydration

[Julio Benavides]

Thanks Ryan. Tissu sections where thin, something about 2 and 3 mm. I 
think they were well fixed (buffered formalin) nut the problem was the 
dehydration. to go from formalin straight into 100% ethanol looks a bit 
too drastic to me.


Thanks for your thoughts

Julio

On 13/02/2015 16:44, Roy, Ryan wrote:

Its well documented in the literature that using graded alcohols in processing 
is advantageous to prevent hardening of the tissue.

How thick are the tissue section cut that are being processed? It is also well 
documented that sections should avoid being cut thicker than 3-4mm as this 
prevents the penetration the fixative as well as the other reagents.

4 hours in each reagent seems excessive... ask other people too since I have 
limited experience.



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Julio Benavides
Sent: Friday, February 13, 2015 10:19 AM
To: histonet@lists.utsouthwestern.edu
Subject: [EXTERNAL] RE: [Histonet] Question regarding dehydration

Hi Ryan,

thanks a lot for your thoughts. These blocks were processed elsewhere and sent 
to us for the cutting and staining. Tissues were dehydrated in five consecutive 
baths of ethanol 100%, 5 hours each (manual processing). Then, they went to 
xilene (three baths, 4 hours each) and paraffin (two batch, 1h30´ each). Truth 
is that I have never seen such processing before. In our lab, with automatic 
processing, we begin with 60% ethanol and go to 70, then 95 and then 100 before 
ethanol/xylene.

I was wondering if anybody has used the 100% ethanol processing before and 
which was the influence over tissues.

Thanks

Julio


 Forwarded Message 
Subject:RE: [EXTERNAL] [Histonet] Question regarding dehydration
Date:   Fri, 13 Feb 2015 09:06:47 -0500
From:   Roy, Ryan ryan@va.gov
To: 'Julio Benavides' j.benavi...@eae.csic.es



If the tissue is fatty it will blow apart on the water bath. If the H and E 
staining is poor its probably the processing.

What is the exact procedure for your process including time that the tissue was 
in each reagent? I would include this in a message to all of histonet as well 
as some may more knowledgeable than myself.

Also, I would recommend using a 95% solution prior to the 100% alcohol (s) if 
not a 70-80% and 95%.


Ryan Roy HTL (ASCP)
Manchester Veterans Affairs Medical Center Manchester New Hampshire

Disclosure: The content of this email does not represent the views or opinons 
of the VA





-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Julio Benavides
Sent: Friday, February 13, 2015 4:26 AM
To: histonet@lists.utsouthwestern.edu
Subject: [EXTERNAL] [Histonet] Question regarding dehydration

Hi there,

we are having problems trying to cut some embedded samples (they crumble in the 
bath and the few cuts we manage to get into HE are crap). These are formalin 
fixed samples (bovine foetal and placenta samples) which went straight from 
formlin into 100º ethanol for the dehydration before clearing. I was wondering 
if such drastic dehydration step (no 60º, 70º or 90º ethanol before the 100º) 
could have damage the tissue. Has anyone have a similar issue before? do you 
think the samples are ruined for histology?

Thanks a lot for your help

Regards

Julio
Instituto de Ganaderia de Montaña
Spain

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RE: [EXTERNAL] RE: [Histonet] Question regarding dehydration

[Elizabeth Chlipala]

Julio

4 to 5 hours a station is way to long for processing samples of this size, 
regardless of what type of tissue or species they are from.  Graduated alcohol 
dehydration is better.  I would process samples of that size (unless it was 
bone, fat or skin) for no longer than 45 - 60 minutes a solution for manual 
processing.  

Here is what a typical processing cycle would look like once the tissue has 
been adequately fixed.

50% alcohol  (you want to start here if you are working with small animal 
tissue such as mice and rats)
70% alcohol 
80% alcohol
95% alcohol
100% alcohol
100% alcohol
Xylene
Xylene
Paraffin
Paraffin
Paraffin

I hope this helps, you may be able to get sections by trimming into the block 
and then soaking them on wet ice for some time (possibly an hour or so).

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 Julio Benavides
Sent: Friday, February 13, 2015 8:43 AM
To: histonet@lists.utsouthwestern.edu
Subject: Re: [EXTERNAL] RE: [Histonet] Question regarding dehydration

Thanks Ryan. Tissu sections where thin, something about 2 and 3 mm. I think 
they were well fixed (buffered formalin) nut the problem was the dehydration. 
to go from formalin straight into 100% ethanol looks a bit too drastic to me.

Thanks for your thoughts

Julio

On 13/02/2015 16:44, Roy, Ryan wrote:
 Its well documented in the literature that using graded alcohols in 
 processing is advantageous to prevent hardening of the tissue.

 How thick are the tissue section cut that are being processed? It is also 
 well documented that sections should avoid being cut thicker than 3-4mm as 
 this prevents the penetration the fixative as well as the other reagents.

 4 hours in each reagent seems excessive... ask other people too since I have 
 limited experience.



 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu 
 [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Julio 
 Benavides
 Sent: Friday, February 13, 2015 10:19 AM
 To: histonet@lists.utsouthwestern.edu
 Subject: [EXTERNAL] RE: [Histonet] Question regarding dehydration

 Hi Ryan,

 thanks a lot for your thoughts. These blocks were processed elsewhere and 
 sent to us for the cutting and staining. Tissues were dehydrated in five 
 consecutive baths of ethanol 100%, 5 hours each (manual processing). Then, 
 they went to xilene (three baths, 4 hours each) and paraffin (two batch, 
 1h30´ each). Truth is that I have never seen such processing before. In our 
 lab, with automatic processing, we begin with 60% ethanol and go to 70, then 
 95 and then 100 before ethanol/xylene.

 I was wondering if anybody has used the 100% ethanol processing before and 
 which was the influence over tissues.

 Thanks

 Julio


  Forwarded Message 
 Subject:  RE: [EXTERNAL] [Histonet] Question regarding dehydration
 Date: Fri, 13 Feb 2015 09:06:47 -0500
 From: Roy, Ryan ryan@va.gov
 To:   'Julio Benavides' j.benavi...@eae.csic.es



 If the tissue is fatty it will blow apart on the water bath. If the H and E 
 staining is poor its probably the processing.

 What is the exact procedure for your process including time that the tissue 
 was in each reagent? I would include this in a message to all of histonet as 
 well as some may more knowledgeable than myself.

 Also, I would recommend using a 95% solution prior to the 100% alcohol (s) if 
 not a 70-80% and 95%.


 Ryan Roy HTL (ASCP)
 Manchester Veterans Affairs Medical Center Manchester New Hampshire

 Disclosure: The content of this email does not represent the views or 
 opinons of the VA





 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu 
 [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Julio 
 Benavides
 Sent: Friday, February 13, 2015 4:26 AM
 To: histonet@lists.utsouthwestern.edu
 Subject: [EXTERNAL] [Histonet] Question regarding dehydration

 Hi there,

 we are having problems trying to cut some embedded samples (they crumble in 
 the bath and the few cuts we manage to get into HE are crap). These are 
 formalin fixed samples (bovine foetal and placenta samples) which went 
 straight from formlin into 100º ethanol for the dehydration before clearing. 
 I was wondering if such drastic dehydration step (no 60º, 70º or 90º ethanol 
 before the 100º) could have damage the tissue. Has anyone have a similar 
 issue before? do you think the samples are ruined for histology?

 Thanks a lot for your help

 Regards

 Julio
 Instituto de Ganaderia de Montaña
 Spain

[Histonet] Question - flow cytometry

[Cartun, Richard]

Does anyone have a protocol to hold specimens for flow cytometry until the 
pathologist examines a slide to confirm that it is 
lymphoproliferative/lymphoma?  We seem to be doing a lot of flow on 
non-lymphoid specimens these days.  Thanks.

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) 972-1596
(860) 545-2204 Fax


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[Histonet] Question?

[Kim Tournear]

All my histonet emails are coming thru my spam folder. Anyone else having this 
problem? And how can I fix it?
Thanks in advance,

Sent from the iPad of Kim Tournear
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Re: [Histonet] Question about high complexity testing for histotechnologists

[Rene J Buesa]

NO, because the license in histotechnology INCLUDES high complexity testing. 
On the other hand there are some tests like FISH with the VYSIS system that 
requires a special training with its own certification.
René J. 
On Tuesday, April 29, 2014 10:15 PM, Delia, Catherine deli...@uhnj.org 
wrote:
  
Does anyone know if there is a regulatory agency that requires high complexity 
testing certification for histotechnologists.

Catherine Susan Delia, BS. HT. ASCP
Chief Technologist, Anatomic Pathology
University Hospital
150 Bergen Street
E-151
Newark, New Jersey 07103
Phone: 973-972-5717
Cell: 908-391-1060
Fax: 973-972-5724
deli...@uhnj.orgmailto:deli...@uhnj.org

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[Histonet] Question about high complexity testing for histotechnologists

[Delia, Catherine]

Does anyone know if there is a regulatory agency that requires high complexity 
testing certification for histotechnologists.

Catherine Susan Delia, BS. HT. ASCP
Chief Technologist, Anatomic Pathology
University Hospital
150 Bergen Street
E-151
Newark, New Jersey 07103
Phone: 973-972-5717
Cell: 908-391-1060
Fax: 973-972-5724
deli...@uhnj.orgmailto:deli...@uhnj.org

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[Histonet] Question for Labs performing Electron Microscopy

[Gauch, Vicki]

Hi everyone,
I have been asked to post the following question regarding EM...
If you are providing EM services in your laboratory, who is performing the EM, 
what is their job title and what are their job duties ( do they process, embed, 
cut,stain,etc. or just certain portions ).
Any information would be greatly appreciated...  Please feel free to e-mail me 
directly or through the Histonet ...

Thanks,
Vicki Gauch
AMCH
Albany, NY


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Re: [Histonet] Question for Labs performing Electron Microscopy

[Rene J Buesa]

We used to do TEM at our lab. The work volume was not high and one of 
histotechnologists was trained to do ALL tasks (fixation → printing the photos 
of the areas selected by the pathologist).
When not doing TEM tasks, he took care of any of any of all HTL tasks for which 
he was also trained.
René J.


On Wed, 3/19/14, Gauch, Vicki gau...@mail.amc.edu wrote:

 Subject: [Histonet] Question for Labs performing Electron Microscopy
 To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu
 Date: Wednesday, March 19, 2014, 10:25 AM
 
 Hi everyone,
 I have been asked to post the following question regarding
 EM...
 If you are providing EM services in your laboratory, who is
 performing the EM, what is their job title and what are
 their job duties ( do they process, embed, cut,stain,etc. or
 just certain portions ).
 Any information would be greatly appreciated...  Please
 feel free to e-mail me directly or through the Histonet ...
 
 Thanks,
 Vicki Gauch
 AMCH
 Albany, NY
 
 
 -
 CONFIDENTIALITY NOTICE: This email and any attachments may
 contain confidential information that is protected by law
 and is for the sole use of the individuals or entities to
 which it is addressed. If you are not the intended
 recipient, please notify the sender by replying to this
 email and destroying all copies of the communication and
 attachments. Further use, disclosure, copying, distribution
 of, or reliance upon the contents of this email and
 attachments is strictly prohibited. To contact Albany
 Medical Center, or for a copy of our privacy practices,
 please visit us on the Internet at www.amc.edu.
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Re: [Histonet] Question for Labs performing Electron Microscopy

[kgrobert]

It's the same for me here.  But then again, we're pure research.

Kathleen

Principal Lab Technician
Neurotoxicology Labs
Molecular Pathology Facility Core
Dept of Pharmacology  Toxicology
Rutgers, the State University of NJ
41 B Gordon Road
Piscataway, NJ 08854
(848) 445-1443


 We used to do TEM at our lab. The work volume was not high and one of
 histotechnologists was trained to do ALL tasks (fixation → printing the
 photos of the areas selected by the pathologist).
 When not doing TEM tasks, he took care of any of any of all HTL tasks for
 which he was also trained.
 René J.

 
 On Wed, 3/19/14, Gauch, Vicki gau...@mail.amc.edu wrote:

  Subject: [Histonet] Question for Labs performing Electron Microscopy
  To: histonet@lists.utsouthwestern.edu
 histonet@lists.utsouthwestern.edu
  Date: Wednesday, March 19, 2014, 10:25 AM

  Hi everyone,
  I have been asked to post the following question regarding
  EM...
  If you are providing EM services in your laboratory, who is
  performing the EM, what is their job title and what are
  their job duties ( do they process, embed, cut,stain,etc. or
  just certain portions ).
  Any information would be greatly appreciated...  Please
  feel free to e-mail me directly or through the Histonet ...

  Thanks,
  Vicki Gauch
  AMCH
  Albany, NY


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  contain confidential information that is protected by law
  and is for the sole use of the individuals or entities to
  which it is addressed. If you are not the intended
  recipient, please notify the sender by replying to this
  email and destroying all copies of the communication and
  attachments. Further use, disclosure, copying, distribution
  of, or reliance upon the contents of this email and
  attachments is strictly prohibited. To contact Albany
  Medical Center, or for a copy of our privacy practices,
  please visit us on the Internet at www.amc.edu.
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[Histonet] Question on Fluorochromes for Histomorphometry

[Herrick, James L. (Jim)]

Dear Histonet Colleagues,

I hope everyone is doing well--

We are going to be using a goat mandible model (embedded in MMA) for a project 
and will need to administer tetracycline and/or calcein (others if suggested) 
for histomorphometric analysis. Would anyone be kind enough to share their 
knowledge on which fluorochromes are optimal, dosing requirements (volume, 
duration and site of injection), etc. We will possibly have three time points. 
The first at 1 month, the second at 3 months and the third at 6 months. Any 
help on this would be greatly appreciated!!

Have a great day!!
Jim



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[Histonet] question about refrigeration

[DiCarlo, Margaret]

Histonetters,

I was wondering how long a bone specimen (portion of pt's tibia in this case) 
can be in a refrigerator before it will decompose and not produce a good 
quality section?  The case was completed at 3:30 pm, refrigerated and then put 
into formalin at 8:15 am the next day?  I am pushing to put specimen into 
formalin immediately after surgery.

Thank you.

Peggy DiCarlo HT (ASCP)
Ortho Bone Lab
Buffalo General Hospital
Buffalo, NY  14203
716-859-1293


The Keeping You Informed section of Kaleida Health`s website features a wealth 
of information, stories and pictures about our valued workforce and the 
tremendous momentum our organization is experiencing. Check us out at: 
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Re: [Histonet] question about refrigeration

[Sean McBride]

Hi Peggy,

In my opinion, it depends upon the objective of the study. Cell autolysis 
begins shortly after death as a result of lack of oxygen supply as well as 
nutrients.  The membranes of the lysosomes break down, and the acid hydrolases 
begin to degrade the cellular 

If you're merely looking to do a macro analysis, such as the quantitation of 
bone, you should be fine. However, if you're hoping to get results at the 
cellular level, either with traditional stains or IHC which relies on intact 
protein structure, you should immerse the specimens  immediately after 
retrieval into formaldehyde solution. Good luck.

Best regards,

Sean

Sean McBride
AFIRM Project Leader
Senior Scientific Specialist
Research Histology Services
Bone Tissue Engineering Center
Carnegie Mellon Research Institute
Suite 4311
700 Technology Drive
Pittsburgh, PA 15219-3124

571-989-BONE (m)
412-268-8275 (o)
412-268-8275 (fax)
smcbr...@andrew.cmu.edu 







On Feb 28, 2014, at 9:12 AM, DiCarlo, Margaret wrote:

 Histonetters,
 
 I was wondering how long a bone specimen (portion of pt's tibia in this case) 
 can be in a refrigerator before it will decompose and not produce a good 
 quality section?  The case was completed at 3:30 pm, refrigerated and then 
 put into formalin at 8:15 am the next day?  I am pushing to put specimen into 
 formalin immediately after surgery.
 
 Thank you.
 
 Peggy DiCarlo HT (ASCP)
 Ortho Bone Lab
 Buffalo General Hospital
 Buffalo, NY  14203
 716-859-1293
 
 
 The Keeping You Informed section of Kaleida Health`s website features a 
 wealth of information, stories and pictures about our valued workforce and 
 the tremendous momentum our organization is experiencing. Check us out at: 
 www.kaleidahealth.org/kyi
 
 CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or 
 previous e-mail messages attached to it are confidential and intended solely 
 for the use of the individual or entity to whom they are addressed. If you 
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 the intended recipient, you are hereby notified that any further review, 
 disclosure, copying, dissemination, distribution, or use of any of the 
 information contained in or attached to this e-mail transmission is strictly 
 prohibited. If you have received this message in error, please notify the 
 sender immediately by e-mail, discard any paper copies, and delete all 
 electronic files of the message. If you are unable to contact the sender or 
 you are not sure as to whether you are the intended recipient, please call 
 Kaleida Health’s Technology Assistance Center at (716) 859-.
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[Histonet] Question about fixation terminology

[Jones, Lynne]

Hello -
Our research group does a fair amount of autoradiography with frozen sections 
and we sometimes perform IHC or routine stains.  I am not a histologist (nor do 
we have one in our current group), so I assumed that the correct answer was 
alcoholic formalin, because the other options were either inappropriate or not 
fixatives.

I was taught (by an old-school cell biologist) that both alcohol and acetone 
act as dehydrating agents when used on frozen sections, and are not true 
fixatives, because they don't cross-link proteins.  Real fixatives don't just 
preserve against decay, they also modify the tissue (i.e., cross-linking or 
denaturing proteins).  I am pretty sure I was taught that acetone does not 
fix tissue, and that fixing tissue to a slide is an imprecise/slang term 
derived from affix.   (This was at least a decade ago, so my memory could be 
faulty.)

How is acetone a fixative?  I thought it just replaced water, and preserved the 
structure of frozen sections by drying them out.  (Please be kind - I'm not a 
histologist.  I've sat at the cryostat sectioning mouse brains, and I know when 
we use it, but I don't understand what the acetone actually does.)

How do you describe the difference between preserving tissue by drying (with 
acetone) and cross-linking the proteins (with alcoholic formalin)?

I would really appreciate clarification from the guru's on HistoNet.  I no 
longer spend much time in the lab, but I edit our manuscripts, so try make sure 
we use the correct terms in describing how the work is done.  (Some reviewers 
get picky about terminology.)

Thanks,

Lynne Jones

Lab Manager
Radiochemistry Research Group
Radiological Chemistry and Imaging Lab
Washington University School of Medicine
St. Louis, MO






Message: 2

Date: Thu, 3 Oct 2013 10:21:42 -0400

From: Lee  Peggy Wenk lpw...@sbcglobal.netmailto:lpw...@sbcglobal.net

Subject: Re: [Histonet] HT HistoDeck question...

To: Watson, Linda linda.wat...@bms.commailto:linda.wat...@bms.com,
  Stephenson, Sheryl

sstephen...@lifecell.commailto:sstephen...@lifecell.com,   
histonet@lists.utsouthwestern.edumailto:histonet@lists.utsouthwestern.edu

Message-ID: 335B9C2DCE414D1CB831DCCB07DEDC94@HP2010

Content-Type: text/plain; format=flowed; charset=iso-8859-1;

reply-type=original



I agree, there is probably more than one correct answer to this question,

depending upon whether you are planning on doing stains for lipids, IHC,

immunofluoresence or muscle enzymes.



But I don't think (A) full strength 37-40% formaldehyde solution would ever

be the correct answer. Unless you put a gauze in the bottom on the coplin

jar, pour a little conc. formaldehyde on the gauze, put a lid on the coplin

jar, and fix the section in formaldehyde vapors. But the question does say

SOLUTION, not VAPOR. So I still think A is wrong. And most likely full

strength acetic acid (C) is wrong - would eat the tissue off the slide.



That leaves cold acetone (B) , which is good for some antibodies and some

enzymes, or alcoholic formalin (D) which might be OK, but most of the time

things either like alcohol and hate formalin, or they like formalin and

don't like alcohol. So I would think most FS that we want to fix would not

particularly like alcoholic formalin.



And none of the solutions listed are good for lipids.



So, given the question (with incomplete information) and the choices of

answers, I would still side with (B) cold acetone.



Now - a little aside - for the questions on the ASCP HT and HTL exams - if

it is a new question, the people on the HT/HTL exam committee would be

looking at it intensely before it goes on the exam for the first time. If

the committee people are having problems answering it (like we are here),

the question would be reworked until all the issues are resolved (such as

putting in for lymph node IHC into the question). If it makes it past the

committee, and the stats from the exam show that many people are having

problems answering it, the question is pulled from the exam and is not used

in the person's score. The question is then sent back to the HT/HTL exam

committee, to try to figure out why examinees were having problems, and the

question reworked again.



As someone who has written exam questions at the school for 20+ years, I can

tell you that it really is hard to write exam questions. I think I've

covered everything in the question so that it is straight forward, and then

half the students read something into the question that I never thought of,

or come up with a written answer that I never considered. So I either have

to throw out the question or give the point to the student, depending upon

what's going on. And then mark the question for a re-write next year.  And

that doesn't include me marking the wrong answer on my master sheet. It

happens!



Peggy A. Wenk, HTL(ASCP)SLS



-Original Message-

From: Watson, Linda

Sent: 

RE: [Histonet] Question about fixation terminology

[joelle weaver]

 Surely I will find those that disagree with this post, however  what I was 
classically trained about fixation categories generally falls within the 
information below...which I took the liberty of  reposting here since it is 
pretty clear  straightforward from Leica. 
http://www.leicabiosystems.com/pathologyleaders/fixation-and-fixatives-1-the-process-of-fixation-and-the-nature-of-fixatives/
 
I hope this will help.
 
 Traditionally fixing agents were termed “coagulant” or “non-coagulant” based 
on their effect on soluble proteins in solution. 7 Coagulant fixatives were 
said to result in a permeable meshwork of protein strands whereas non-coagulant 
fixatives which are additive in nature, formed extensive cross-links producing 
a less permeable gel. These terms are still encountered in modern histological 
literature but a more systematic approach has recently been taken to 
classification. 2  There are two major mechanisms which are important in 
fixation of proteins and protein complexes: denaturation, and addition and 
cross-link formation.  Denaturation:  Most commonly this effect is induced by 
dehydrants such as the alcohols or acetone. These reagents remove and replace 
free water in cells and tissues and cause a change in the tertiary structure of 
proteins by destabilizing hydrophobic bonding. Hydrophobic areas, frequently 
found on the inside of protein molecules, are released from the repulsion of 
water and become free to occupy a greater area.  hydrophilic areas of protein 
water molecules are loosely bound by hydrogen bonds and removal of water also 
destabilizes these bonds. The changes produced in the conformation of the 
protein molecules cause a change in the solubility of the protein, rendering 
water soluble proteins insoluble, a change that is largely irreversible if the 
protein is returned to an aqueous environment.  2, 7  Addition and cross-link 
formation:  The non-coagulant fixing agents chemically react with proteins and 
other cell and tissue components, becoming bound to them by addition and 
forming inter-molecular and intra-molecular cross-links. Because these agents 
are reactive compounds they bind to a variety of chemical groups in tissues, 
often affecting the charge at the site of attachment. This can have an effect 
on the subsequent staining characteristics of a particular protein as well as 
altering its molecular conformation and thus its solubility . For further and 
more in depth discussion of formalin fixation ( and the article I am usually 
referred to in order to make corrections to my own chemistry in publications) 
is  1985!! Fox, et al. Formaldehyde Fixation,  Journal of Histochemistry and 
Cytochemistry, vol. 33, No. 8. pp. 845-855. pdf available at 
http://www.gofindpdf.com/readpdf/formaldehyde-fixation.html  Joelle Weaver 
MAOM, HTL (ASCP) QIHC
  From: jone...@mir.wustl.edu
 To: histonet@lists.utsouthwestern.edu
 Date: Thu, 3 Oct 2013 19:32:51 +
 CC: 
 Subject: [Histonet] Question about fixation terminology
 
 Hello -
 Our research group does a fair amount of autoradiography with frozen sections 
 and we sometimes perform IHC or routine stains.  I am not a histologist (nor 
 do we have one in our current group), so I assumed that the correct answer 
 was alcoholic formalin, because the other options were either inappropriate 
 or not fixatives.
 
 I was taught (by an old-school cell biologist) that both alcohol and acetone 
 act as dehydrating agents when used on frozen sections, and are not true 
 fixatives, because they don't cross-link proteins.  Real fixatives don't 
 just preserve against decay, they also modify the tissue (i.e., cross-linking 
 or denaturing proteins).  I am pretty sure I was taught that acetone does not 
 fix tissue, and that fixing tissue to a slide is an imprecise/slang term 
 derived from affix.   (This was at least a decade ago, so my memory could 
 be faulty.)
 
 How is acetone a fixative?  I thought it just replaced water, and preserved 
 the structure of frozen sections by drying them out.  (Please be kind - I'm 
 not a histologist.  I've sat at the cryostat sectioning mouse brains, and I 
 know when we use it, but I don't understand what the acetone actually does.)
 
 How do you describe the difference between preserving tissue by drying (with 
 acetone) and cross-linking the proteins (with alcoholic formalin)?
 
 I would really appreciate clarification from the guru's on HistoNet.  I no 
 longer spend much time in the lab, but I edit our manuscripts, so try make 
 sure we use the correct terms in describing how the work is done.  (Some 
 reviewers get picky about terminology.)
 
 Thanks,
 
 Lynne Jones
 
 Lab Manager
 Radiochemistry Research Group
 Radiological Chemistry and Imaging Lab
 Washington University School of Medicine
 St. Louis, MO
 
 
 
 
 
 
 Message: 2
 
 Date: Thu, 3 Oct 2013 10:21:42 -0400
 
 From: Lee  Peggy Wenk lpw...@sbcglobal.netmailto:lpw...@sbcglobal.net
 
 Subject: Re: [Histonet] HT

[Histonet] Question for Our IHC Area

[Pam Marcum]

Is anyone having an issue with Ventana's Pan cytokeratin?  Please answer 
offline. 

  

Pam Marcum 









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RE: [Histonet] Question - OR specimens to Pathology

[Horn, Hazel V]

Luckily our Frozen Section Lab is in the OR.

Hazel Horn
Supervisor of Histology/Autopsy/Transcription
Anatomic Pathology
Arkansas Children's Hospital
1 Children's Way | Slot 820| Little Rock, AR 72202
501.364.4240 direct | 501.364.1241 fax
hor...@archildrens.org
archildrens.org






-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Grantham, 
Andrea L - (algranth)
Sent: Thursday, August 01, 2013 4:52 PM
Cc: HISTONET
Subject: Re: [Histonet] Question - OR specimens to Pathology

OR - you can get a rubbermaid took box and slap a few biohazard stickers on it 
and put some formalin pads inside and you have a transport box for surgical 
specimens.
I happen to know that that is how this box was invented.




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

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





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[Histonet] Question - OR specimens to Pathology

[Richard Cartun]

How are other hospitals transporting fresh tissue specimens from operating 
rooms to the Pathology Frozen Section room for an intraoperative consultation?  
Occasionally, we get specimens delivered on towels and I don't think this is 
appropriate.  I think they should be in a container that can be sealed along 
with the appropriate patient identification.  Thank you.
 
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
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RE: [Histonet] Question - OR specimens to Pathology

[Elizabeth Chlipala]

Richard

MOPEC has an item called pathport it's about the size of a tool box that what 
we used to use to carry frozen sections from the surgery to pathology.

http://www.mopec.com/product/1829/pathport/

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 Richard Cartun 
[rcar...@harthosp.org]
Sent: Thursday, August 01, 2013 2:26 PM
To: Histonet
Subject: [Histonet] Question - OR specimens to Pathology

How are other hospitals transporting fresh tissue specimens from operating 
rooms to the Pathology Frozen Section room for an intraoperative consultation?  
Occasionally, we get specimens delivered on towels and I don't think this is 
appropriate.  I think they should be in a container that can be sealed along 
with the appropriate patient identification.  Thank you.

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

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Re: [Histonet] Question - OR specimens to Pathology

[Grantham, Andrea L - (algranth)]

OR - you can get a rubbermaid took box and slap a few biohazard stickers on it 
and put some formalin pads inside and you have a transport box for surgical 
specimens.
I happen to know that that is how this box was invented.




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

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





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Re: [Histonet] Question - OR specimens to Pathology

[Heather D'orazio]

I wouldn't recommend putting fresh tissue specimens destined for frozen tissue 
processing into or on formalin before it is grossed, etc. Our Mohs surgeon 
walks the surgical specimen from the OR directly to the Mohs lab on an oriented 
piece of gauze in a clean petri dish along with map, chart, etc. Surgeon and 
tech then have the opportunity to discuss specimen before tech processes. This 
process passes muster for CLIA and works well for us.

Heather D'Orazio
Mohs Tech
Rogers Dermatology Clinic
1727 West College
Bozeman MT 59715
(406) 587-4432
 


 From: Grantham, Andrea L - (algranth) algra...@email.arizona.edu
To: 
Cc: HISTONET histonet@lists.utsouthwestern.edu 
Sent: Thursday, August 1, 2013 3:51 PM
Subject: Re: [Histonet] Question - OR specimens to Pathology
  

OR - you can get a rubbermaid took box and slap a few biohazard stickers on it 
and put some formalin pads inside and you have a transport box for surgical 
specimens.
I happen to know that that is how this box was invented.




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

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





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[Histonet] Question - MDM

[Richard Cartun]

Is anyone using a commercially available antibody (not RUO) for the 
immunohistochemical detection of MDM in liposarcoma?  Thank you.
 
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
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[Histonet] Question regarding appropriate use of histonet

[Henry Chidgey]

Howdy Folks,

As I am sure you can tell we are subscribers to histonet and appreciate the
generosity of members in helping solve the occasional problem we encounter.
We are in need of finding either an individual or lab in our area to perform
mammal root sectioning and slide preparation for the forensic mammal aging
business we operate located near Burnet, Texas in the Texas Hill Country. I
would like to reach out to your subscriber base to see if anyone is
interested in doing some part time work for us or know someone who might be.
Please let me know if that would be an appropriate use of the site.

Thanks,

Henry Chidgey

CoFounder

 

See what folks are saying about us http://www.deerage.com/testimonials.htm


Wildlife Analytical Laboratories

 www.DeerAge.com http://www.deerage.com/ 

 PO Box 295

 1303 CR 118 B

 Burnet, Texas 78611

(512) 756-1989

(512) 663-9476 Cell

Serving Wildlife Stewards, Ranchers  Hunters Worldwide

Customer Service and Quality of Analysis are our Priority and Passion

Facebook http://www.facebook.com/DeerAge   Eagle Soybeans
http://deerage.com/eagle_seed/ 

 

 

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Re: [Histonet] question

[Alan Bright]

I do not see that as UV will not decontaminate areas and tissue that are not in 
the UV light path or thick tissue.
Alan Bright

Sent from my iPhone

On 20 Mar 2013, at 23:38, joelle weaver joellewea...@hotmail.com wrote:

 Maybe that is why some like the UV decontamination option? I know I prefer 
 it.  If there is aerolizing of any material or particles after that cycle, at 
 least it whatever was trapped in the chamber would  have been decontaminated 
 when you then vaccum. I think that the UV models have their own vaccum as I 
 recall.  Plus it is MUCH faster than the old defrots, wipe out, vaccum and 
 rinse steps and air dry steps needed to decontaminate some ( usually older) 
 cryostats.
 
 
 
 Joelle Weaver MAOM, HTL (ASCP) QIHC
  
  From: abri...@brightinstruments.com
  Date: Wed, 20 Mar 2013 21:37:45 +
  To: marilyn.a.we...@kp.org
  Subject: Re: [Histonet] question
  CC: histonet@lists.utsouthwestern.edu
  
  Strange about the gloves when vacuuming out fresh tissue trimmings from 
  cryostats and exhausting the air back into the lab is done.
  
  Alan Bright
  
  On 20 Mar 2013, at 19:07, marilyn.a.we...@kp.org marilyn.a.we...@kp.org 
  wrote:
  
   Thank you for your response about the squames and we are all wearing 
   gloves now, if one uses them then we all do is my motto, although in 50 
   years of cutting, this is the first time this has come up. . guess I was 
   lucky. anyway, the same pathologists would like to know if there is a 
   standard about wearing gloves in the histo world and if so, is it because 
   of squames or although the tissue is fixed, is there chances of 
   infections 
   or something else. thank you.
   
   NOTICE TO RECIPIENT: If you are not the intended recipient of this 
   e-mail, you are prohibited from sharing, copying, or otherwise using or 
   disclosing its contents.  If you have received this e-mail in error, 
   please notify the sender immediately by reply e-mail and permanently 
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 Spam
 Not spam
 Forget previous vote
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RE: [Histonet] question

[joelle weaver]

I guess I didn't understand what debris you were speaking of, yes it would only 
decontaminate what is within the chamber. I guess I never had much issue with 
any blowing




Joelle Weaver MAOM, HTL (ASCP) QIHC
 CC: marilyn.a.we...@kp.org; histonet@lists.utsouthwestern.edu
From: abri...@brightinstruments.com
Subject: Re: [Histonet] question
Date: Thu, 21 Mar 2013 10:25:41 +
To: joellewea...@hotmail.com

I do not see that as UV will not decontaminate areas and tissue that are not in 
the UV light path or thick tissue.Alan Bright

Sent from my iPhone
On 20 Mar 2013, at 23:38, joelle weaver joellewea...@hotmail.com wrote:




Maybe that is why some like the UV decontamination option? I know I prefer it.  
If there is aerolizing of any material or particles after that cycle, at least 
it whatever was trapped in the chamber would  have been decontaminated when you 
then vaccum. I think that the UV models have their own vaccum as I recall.  
Plus it is MUCH faster than the old defrots, wipe out, vaccum and rinse steps 
and air dry steps needed to decontaminate some ( usually older) cryostats.





Joelle Weaver MAOM, HTL (ASCP) QIHC
 
 From: abri...@brightinstruments.com
 Date: Wed, 20 Mar 2013 21:37:45 +
 To: marilyn.a.we...@kp.org
 Subject: Re: [Histonet] question
 CC: histonet@lists.utsouthwestern.edu
 
 Strange about the gloves when vacuuming out fresh tissue trimmings from 
 cryostats and exhausting the air back into the lab is done.
 
 Alan Bright
 
 On 20 Mar 2013, at 19:07, marilyn.a.we...@kp.org marilyn.a.we...@kp.org 
 wrote:
 
  Thank you for your response about the squames and we are all wearing 
  gloves now, if one uses them then we all do is my motto, although in 50 
  years of cutting, this is the first time this has come up. . guess I was 
  lucky. anyway, the same pathologists would like to know if there is a 
  standard about wearing gloves in the histo world and if so, is it because 
  of squames or although the tissue is fixed, is there chances of infections 
  or something else. thank you.
  
  NOTICE TO RECIPIENT:  If you are not the intended recipient of this 
  e-mail, you are prohibited from sharing, copying, or otherwise using or 
  disclosing its contents.  If you have received this e-mail in error, 
  please notify the sender immediately by reply e-mail and permanently 
  delete this e-mail and any attachments without reading, forwarding or 
  saving them.  Thank you.
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[Histonet] question

[Marilyn . A . Weiss]

Thank you for your response about the squames and we are all wearing 
gloves now, if one uses them then we all do is my motto, although in 50 
years of cutting, this is the first time this has come up. . guess I was 
lucky. anyway, the same pathologists would like to know if there is a 
standard about wearing gloves in the histo world and if so, is it because 
of squames or although the tissue is fixed, is there chances of infections 
or something else. thank you.

NOTICE TO RECIPIENT:  If you are not the intended recipient of this 
e-mail, you are prohibited from sharing, copying, or otherwise using or 
disclosing its contents.  If you have received this e-mail in error, 
please notify the sender immediately by reply e-mail and permanently 
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Re: [Histonet] question

[Alan Bright]

Strange about the gloves when vacuuming out fresh tissue trimmings from 
cryostats and exhausting the air back into the lab is done.

Alan Bright

On 20 Mar 2013, at 19:07, marilyn.a.we...@kp.org marilyn.a.we...@kp.org 
wrote:

 Thank you for your response about the squames and we are all wearing 
 gloves now, if one uses them then we all do is my motto, although in 50 
 years of cutting, this is the first time this has come up. . guess I was 
 lucky. anyway, the same pathologists would like to know if there is a 
 standard about wearing gloves in the histo world and if so, is it because 
 of squames or although the tissue is fixed, is there chances of infections 
 or something else. thank you.
 
 NOTICE TO RECIPIENT:  If you are not the intended recipient of this 
 e-mail, you are prohibited from sharing, copying, or otherwise using or 
 disclosing its contents.  If you have received this e-mail in error, 
 please notify the sender immediately by reply e-mail and permanently 
 delete this e-mail and any attachments without reading, forwarding or 
 saving them.  Thank you.
 ___
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 Histonet@lists.utsouthwestern.edu
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 -- 
 

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RE: [Histonet] question

[joelle weaver]

Maybe that is why some like the UV decontamination option? I know I prefer it.  
If there is aerolizing of any material or particles after that cycle, at least 
it whatever was trapped in the chamber would  have been decontaminated when you 
then vaccum. I think that the UV models have their own vaccum as I recall.  
Plus it is MUCH faster than the old defrots, wipe out, vaccum and rinse steps 
and air dry steps needed to decontaminate some ( usually older) cryostats.




Joelle Weaver MAOM, HTL (ASCP) QIHC
  From: abri...@brightinstruments.com
 Date: Wed, 20 Mar 2013 21:37:45 +
 To: marilyn.a.we...@kp.org
 Subject: Re: [Histonet] question
 CC: histonet@lists.utsouthwestern.edu
 
 Strange about the gloves when vacuuming out fresh tissue trimmings from 
 cryostats and exhausting the air back into the lab is done.
 
 Alan Bright
 
 On 20 Mar 2013, at 19:07, marilyn.a.we...@kp.org marilyn.a.we...@kp.org 
 wrote:
 
  Thank you for your response about the squames and we are all wearing 
  gloves now, if one uses them then we all do is my motto, although in 50 
  years of cutting, this is the first time this has come up. . guess I was 
  lucky. anyway, the same pathologists would like to know if there is a 
  standard about wearing gloves in the histo world and if so, is it because 
  of squames or although the tissue is fixed, is there chances of infections 
  or something else. thank you.
  
  NOTICE TO RECIPIENT:  If you are not the intended recipient of this 
  e-mail, you are prohibited from sharing, copying, or otherwise using or 
  disclosing its contents.  If you have received this e-mail in error, 
  please notify the sender immediately by reply e-mail and permanently 
  delete this e-mail and any attachments without reading, forwarding or 
  saving them.  Thank you.
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RE: [Histonet] question

[joelle weaver]

sorry for the misspelling in my previous post.




Joelle Weaver MAOM, HTL (ASCP) QIHC
  From: joellewea...@hotmail.com
 To: abri...@brightinstruments.com; marilyn.a.we...@kp.org
 Date: Wed, 20 Mar 2013 23:38:04 +
 Subject: RE: [Histonet] question
 CC: histonet@lists.utsouthwestern.edu
 
 Maybe that is why some like the UV decontamination option? I know I prefer 
 it.  If there is aerolizing of any material or particles after that cycle, at 
 least it whatever was trapped in the chamber would  have been decontaminated 
 when you then vaccum. I think that the UV models have their own vaccum as I 
 recall.  Plus it is MUCH faster than the old defrots, wipe out, vaccum and 
 rinse steps and air dry steps needed to decontaminate some ( usually older) 
 cryostats.
 
 
 
 
 Joelle Weaver MAOM, HTL (ASCP) QIHC
   From: abri...@brightinstruments.com
  Date: Wed, 20 Mar 2013 21:37:45 +
  To: marilyn.a.we...@kp.org
  Subject: Re: [Histonet] question
  CC: histonet@lists.utsouthwestern.edu
  
  Strange about the gloves when vacuuming out fresh tissue trimmings from 
  cryostats and exhausting the air back into the lab is done.
  
  Alan Bright
  
  On 20 Mar 2013, at 19:07, marilyn.a.we...@kp.org marilyn.a.we...@kp.org 
  wrote:
  
   Thank you for your response about the squames and we are all wearing 
   gloves now, if one uses them then we all do is my motto, although in 50 
   years of cutting, this is the first time this has come up. . guess I was 
   lucky. anyway, the same pathologists would like to know if there is a 
   standard about wearing gloves in the histo world and if so, is it because 
   of squames or although the tissue is fixed, is there chances of 
   infections 
   or something else. thank you.
   
   NOTICE TO RECIPIENT:  If you are not the intended recipient of this 
   e-mail, you are prohibited from sharing, copying, or otherwise using or 
   disclosing its contents.  If you have received this e-mail in error, 
   please notify the sender immediately by reply e-mail and permanently 
   delete this e-mail and any attachments without reading, forwarding or 
   saving them.  Thank you.
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[Histonet] Question on Estrogen antibodies for fish

[Shirley A. Powell]

I am asking this for a researcher.  Please respond to her at 
klar_elizab...@columbusstate.edumailto:klar_elizab...@columbusstate.edu. Her 
name is Ely Klar.

She is looking for any form of estrogen antibody that works on bass fish 
gonads.  Vendor's responses welcome.


Shirley A. Powell, HT(ASCP)HTL, QIHC
Technical Director
Histology Curricular Support Laboratory
Mercer University School of Medicine
1550 College Street
Macon, GA  31207
478-301-2374 Lab
478-301-5489 Fax

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[Histonet] Question for Alkaline Phosphatase / Biotin System Users

[Amy Porter]

We are using standard avidin / biotin staining with alkaline phosphatase
labeling and sometimes, not every time and not every slide in the same run
we get residual blotchy spots and streaking of the reaction on and off the
tissue.  We have increased to two rinses after primary and two rinses after
alk phos reagent prior to substrate; still happening on some slides...anyone
else have this experience or type of problem?  Any experiences or
suggestions are appreciated.  Just can't figure out what is causing it..

 

Amy S. Porter, HT(ASCP) QIHC

Michigan State University

Investigative HistoPathology Laboratory

port...@msu.edu

 

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[Histonet] Question about sectioning brain tissue at 10-15 microns

[Thurby, Christina]

Hi,
Can anyone with experience in cutting thick FFPE brain sections please give 
advice on how to get a thick brain section without it curling up before it gets 
to the waterbath?  We are doing okay at cutting these at 10 microns, but are 
trying to get some 12 micron sections and the section just curls as it is being 
cut on the microtome.We use Paraplast - just in case anyone needs to know.  
I did read that having a lower temp melting point paraffin sometimes helps with 
this issue.  Any other thoughts from folks that routinely do this?
Thanks!
Christina Thurby
Bristol Myers Squibb
812-307-2093




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[Histonet] Question about sectioning brain tissue at 10-15 microns

[Tim Wheelock]

Hi Christina:

I use Paraplast to infiltrate the formalin fixed brain tissue, but I use 
Surgipath Embedding Medium from Leica (Catalogue number 3801320)  to 
embed the tissue.

(Surgipath does have about the same melting point as the Paraplast)
Even the 10 micron sections (for the Congo red stain) tend to curl, but 
I just guide them off of the disposable blade with a brush or a pick.


Hope this helps,

Tim

Tim Wheelock
Harvard Brain Bank
McLean Hospital
Belmont MA
617-855-3592

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[Histonet] RE: Histonet Question

[Tyrrell, Jannifer]

I'm new to the Histonet website, but not new to histotechnology.

I'm trying to salvage some irreplaceable Zucker Diabetic Fatty rat tissue 
samples that were embedded in paraffin in May 2009. They have sat on my bench 
since they were embedded as we thought the entire experiment was a huge 
failure.  However, recent examination of some of the flash-frozen tissues has 
shown us that the experiment was actually a huge success.

Which leads me to my issues and question...

I needed to section these tissues (liver  pancreas) and have attempted to do 
so by several troubleshooting means - soaking the block in warm water then 
rapid cooling, place ice on the block then rapid slicing, changing the blade, 
using a different microtome, changing the blade angle, you name it, I think 
I've tried it.

Now I've got a bunch of sections that look like accordions that will not 
flatten, but hardly anything left in the actual blocks. These horrible sections 
are on slides, but not stained yet.  I only put them on slides because I NEED 
the tissues. But to stain the sections as they are would be a waste of the 
tissues and the reagents.

I saw a protocol for formol-glycerol re-hydration and have thought about using 
it...but now I can't get the tissue sections off of the glass slides.

ANY and all help or advice would be INCREDIBLY and GREATLY appreciated! This is 
the last bit of work I have to complete in my dissertation project and the 
data, I'm certain, will be astounding.

Jannifer Tyrrell
NIEHS/NIH-Sponsored Pre-Doctoral
Ruth Kirschstein Federal Grant Recipient
PhD Candidate
Department of Pathology
Wayne State University
School of Medicine
550 E. Canfield Ave.
111 Lande Building
Detroit, MI 48201

jtyrr...@med.wayne.edu
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[Histonet] Question about slide drying/ convection ovens

[Natalie Nagy]

Hi everyone,
  Just have a quick question for all of you out there in 
histo-land. Have you ever bought or worked with a gravity convection oven (the 
thermo shandon 20GC), for drying of slides?? If so..how was it? Did it properly 
dry all of your slides, did it keep temp?? I am thinking about buying one, and 
wanted some opinions. Or do you all prefer a real forced air slide drying oven?
Thanks in advance,
   
  Natalie Nagy (HT)ASCP
Histology Supervisor
Holyoke Medical Center
Holyoke, MA.
 


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Re: [Histonet] Question about slide drying/ convection ovens

[Rene J Buesa]

Natalia: 
I just cannot understand the concept of gravity convection oven because if 
you say gravity by definition that will mean that the heat will work by 
gravity and any heated air goes, also by definition, against gravity because 
any heat air will go UP and not down, hence the hot air balloons.
Probably the title is just a catchy but wrongly selected sales mimic, 
something new and different.
Other than that, and going to your question, no I have never used such an oven.
Ovens are of the convection type and in all of them the heating elements are at 
the bottom and the heated air goes UP.
René J.

From: Natalie Nagy nagy_nata...@holyokehealth.com
To: histonet@lists.utsouthwestern.edu 
Sent: Monday, January 28, 2013 12:40 PM
Subject: [Histonet] Question about slide drying/ convection ovens

Hi everyone,
                  Just have a quick question for all of you out there in 
histo-land. Have you ever bought or worked with a gravity convection oven (the 
thermo shandon 20GC), for drying of slides?? If so..how was it? Did it properly 
dry all of your slides, did it keep temp?? I am thinking about buying one, and 
wanted some opinions. Or do you all prefer a real forced air slide drying oven?
Thanks in advance,
              
      Natalie Nagy (HT)ASCP
Histology Supervisor
Holyoke Medical Center
Holyoke, MA.



CONFIDENTIALITY NOTICE: This email communication and any attachments may 
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review, disclosure, dissemination, distribution or copying of it or its 
contents is prohibited. If you have received this communication in error, 
please reply to the sender immediately and destroy all copies of this 
communication and any attachments. For further information regarding Holyoke 
Medical Center's privacy policy, Please visit our Internet web site at 
http://www.holyokehealth.com
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RE: [Histonet] Question about slide drying/ convection ovens

[Morken, Timothy]

There is a reason to distinguish the type of oven but the terms are used 
loosely and so cause confusion.

Convection is used to describe in nature to describe the natural circulation 
of heat and can be of natural gravity convection in which heat rises, or 
forced in which some kind of external force spreads the heat faster Ie, wind.

Strictly speaking a Convection Oven is one that uses a fan to distribute the 
heat. That is opposed to Conventional Ovens that do not use fans and rely on 
gravity to distribute heat (heat rises, setting up convection currents).

However, if you look at equipment catalogs you will see terms like Gravity 
Convection and Forced Air to distinguish between ovens without and with fans 
respectively. 

Most ovens in use are gravity-convection ovens that rely on the natural rise 
of heated air to circulate the heat. They don' t use fans.

In contrast is the forced-air convection oven that uses a fan to circulate air 
and ensure even heating throughout the oven. Forced air can heat/dry a sample 
faster if the fan is set to circulate air at a faster rate than would happen by 
natural convection.

So, the moral of the story is to read the description of the oven to find out 
exactly what kind of oven it really is!


Tim Morken
UCSF Pathology


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Monday, January 28, 2013 10:06 AM
To: Natalie Nagy; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Question about slide drying/ convection ovens

Natalia: 
I just cannot understand the concept of gravity convection oven because if 
you say gravity by definition that will mean that the heat will work by 
gravity and any heated air goes, also by definition, against gravity because 
any heat air will go UP and not down, hence the hot air balloons.
Probably the title is just a catchy but wrongly selected sales mimic, 
something new and different.
Other than that, and going to your question, no I have never used such an oven.
Ovens are of the convection type and in all of them the heating elements are at 
the bottom and the heated air goes UP.
René J.

From: Natalie Nagy nagy_nata...@holyokehealth.com
To: histonet@lists.utsouthwestern.edu
Sent: Monday, January 28, 2013 12:40 PM
Subject: [Histonet] Question about slide drying/ convection ovens

Hi everyone,
                  Just have a quick question for all of you out there in 
histo-land. Have you ever bought or worked with a gravity convection oven (the 
thermo shandon 20GC), for drying of slides?? If so..how was it? Did it properly 
dry all of your slides, did it keep temp?? I am thinking about buying one, and 
wanted some opinions. Or do you all prefer a real forced air slide drying oven?
Thanks in advance,
              
      Natalie Nagy (HT)ASCP
Histology Supervisor
Holyoke Medical Center
Holyoke, MA.



CONFIDENTIALITY NOTICE: This email communication and any attachments may 
contain confidential and privileged information for the use of the designated 
recipients named above. If you are not the intended recipient, you are hereby 
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review, disclosure, dissemination, distribution or copying of it or its 
contents is prohibited. If you have received this communication in error, 
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[Histonet] Question

[Courtney Pierce]

Are IHC high complexity test.

Courtney Pierce
IHC Specialist
Quintiles
Translational RD - Oncology
Innovation
Navigating the new health

777 Oakmont Lane Suite 100
Westmont,IL 60559

Office: + 630-203-6234
courtney.pie...@quintiles.com

clinical | commercial | consulting | capital


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This electronic message, including its attachments, is COMPANY CONFIDENTIAL
and may contain PROPRIETARY or LEGALLY PRIVILEGED information.  If you are 
not the intended recipient, you are hereby notified that any use, disclosure,
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message in error, please immediately notify the sender by reply e-mail and
permanently delete this message and its attachments, along with any copies
thereof. Thank you. 


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Re: [Histonet] Question

[Rene J Buesa]

Yes!
René J.

From: Courtney Pierce courtney.pie...@quintiles.com
To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu 
Sent: Tuesday, January 22, 2013 1:04 PM
Subject: [Histonet] Question

Are IHC high complexity test.

Courtney Pierce
IHC Specialist
Quintiles
Translational RD - Oncology
Innovation
Navigating the new health

777 Oakmont Lane Suite 100
Westmont,IL 60559

Office: + 630-203-6234
courtney.pie...@quintiles.com

clinical | commercial | consulting | capital


**  IMPORTANT--PLEASE READ  
This electronic message, including its attachments, is COMPANY CONFIDENTIAL
and may contain PROPRIETARY or LEGALLY PRIVILEGED information.  If you are 
not the intended recipient, you are hereby notified that any use, disclosure,
copying, or distribution of this message or any of the information included
in it is unauthorized and strictly prohibited.  If you have received this
message in error, please immediately notify the sender by reply e-mail and
permanently delete this message and its attachments, along with any copies
thereof. Thank you. 


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Re: [Histonet] Question

[Mark Tarango]

The staining portion is not high complexity.  The reading of the slide is.

On Tue, Jan 22, 2013 at 10:04 AM, Courtney Pierce 
courtney.pie...@quintiles.com wrote:

 Are IHC high complexity test.

 Courtney Pierce
 IHC Specialist
 Quintiles
 Translational RD - Oncology
 Innovation
 Navigating the new health

 777 Oakmont Lane Suite 100
 Westmont,IL 60559

 Office: + 630-203-6234
 courtney.pie...@quintiles.com

 clinical | commercial | consulting | capital


 **  IMPORTANT--PLEASE READ  
 This electronic message, including its attachments, is COMPANY CONFIDENTIAL
 and may contain PROPRIETARY or LEGALLY PRIVILEGED information.  If you are
 not the intended recipient, you are hereby notified that any use,
 disclosure,
 copying, or distribution of this message or any of the information included
 in it is unauthorized and strictly prohibited.  If you have received this
 message in error, please immediately notify the sender by reply e-mail and
 permanently delete this message and its attachments, along with any copies
 thereof. Thank you.
 

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[Histonet] Question removal

[Chris Winans]

Hello,
 How do I go about removing a question from this site? Thanks!
Chris

ENFD Supervisor
Bako Pathology Services
ch...@bakopathology.commailto:ch...@bakopathology.com


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Re: [Histonet] Question removal

[Rene J Buesa]

As in any e-mail, once you press send, you cannot withdraw/remove the message.
You may try to write to the master of the site.
René J.

From: Chris Winans ch...@bakopathology.com
To: Histonet@lists.utsouthwestern.edu Histonet@lists.utsouthwestern.edu 
Sent: Monday, January 21, 2013 7:59 AM
Subject: [Histonet] Question removal

Hello,
How do I go about removing a question from this site? Thanks!
Chris

ENFD Supervisor
Bako Pathology Services
ch...@bakopathology.commailto:ch...@bakopathology.com


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[Histonet] question(s)

[Webb, Dorothy L]

How does everyone handle storing extra paraffin sections that are cut as a 
standard on certain protocols, such as prostate needle bx's?  We currently 
place them on a slide and save until the case is signed out.  I am concerned 
with the amount of waste and cost with the way we are doing it and would like 
other ideas.  If you keep the ribbons laid out on paper, do you stack the 
sheets of paper and is that problematic when it comes time to need that certain 
cut for IHC?

Also, please let me know of those who have the VIP 6, how you like it, pros and 
cons and also the Leica ASP300S.
Would greatly appreciate all feedback on both requests and thank you so very 
much!!

Dorothy Webb , HT(ASCP)
Regions Histology Technical Specialist



  
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Re: [Histonet] question(s)

[Rene J Buesa]

The sectioning protocols/sequences are determined by the pathologists and what 
they think is required to make diagnoses. We also stored the slides with extras 
sections until the case is signed and get an additional OK from the 
signing pathologist before disposing of the unused sections.
I would never ever place ribbons of sections on paper. That is calling for 
disaster in the event that some sections are required.
If you think there is too much extra work and materials wastes your only 
solution is to ask the pathologists to review their requirements as to extra 
sections, specially those the pathologist may want to use for IHC. In some 
protocols the pathologists require automated IHC procedures before signing the 
case. All these issues are to be resolved by the pathologists. If the excess 
work is too much, you can always talk with your manager to get help regarding 
the protocols.
René J.

From: Webb, Dorothy L dorothy.l.w...@healthpartners.com
To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu 
Sent: Friday, December 7, 2012 1:10 PM
Subject: [Histonet] question(s)

How does everyone handle storing extra paraffin sections that are cut as a 
standard on certain protocols, such as prostate needle bx's?  We currently 
place them on a slide and save until the case is signed out.  I am concerned 
with the amount of waste and cost with the way we are doing it and would like 
other ideas.  If you keep the ribbons laid out on paper, do you stack the 
sheets of paper and is that problematic when it comes time to need that certain 
cut for IHC?

Also, please let me know of those who have the VIP 6, how you like it, pros and 
cons and also the Leica ASP300S.
Would greatly appreciate all feedback on both requests and thank you so very 
much!!

Dorothy Webb , HT(ASCP)
Regions Histology Technical Specialist



  
This e-mail and any files transmitted with it are confidential and are intended 
solely for the use of the individual or entity to whom they are addressed. If 
you are not the intended recipient or the individual responsible for delivering 
the e-mail to the intended recipient, please be advised that you have received 
this e-mail in error and that any use, dissemination, forwarding, printing, or 
copying of this e-mail is strictly prohibited.

If you have received this communication in error, please return it to the 
sender immediately and delete the original message and any copy of it from your 
computer system. If you have any questions concerning this message, please 
contact the sender. Disclaimer R001.0
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[Histonet] Question on frozen section IHC and tissue adhesion

[Thurby, Christina]

Hi all,
I am performing IHC on tissue that was formalin fixed for 48 hours and then 
embedded in OCT.  I have been using Gold Plus slides from Thermo Fisher.I 
am staining rat spleen and mandibular lymph nodes.  With a peroxidase method, 
the tissue completely digests off the slides as soon as I apply the peroxidase 
block (looks like pouring hydrogen peroxide on a cut - just turns white and 
bubbles off of the slide).  I have tried various concentrations of peroxidase 
block with no luck.  With the alkaline phos method, the tissue is staying 
adhered to the slide, but I am getting significant 'tissue lifting' which makes 
the pathologists interpretation very difficult.  Is there anyone out there with 
experience in doing IHC on formalin fixed frozen sections that has experienced 
similar 'tissue lifting' problems?  I have ran this antibody on this tissue 
with immunofluorescence successfully - that may be the only choice we have for 
this particular tissue.  Would really like to be able to perform this staining 
with either an HRP or AP method for light microscopy if possible.

Christina Thurby
Bristol Myers Squibb
812-307-2093





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Re: [Histonet] Question on frozen section IHC and tissue adhesion

[Rene J Buesa]

Formalin fixed tissue is not adequate for frozen sectioning unless you place it 
in sucrose first before going into OCT and frozen sectioning. Spleen is 
particularly difficult for its high blood contents. Ideally you should try to 
obtain unfixed tissue.
René J.



From: Thurby, Christina christina.thu...@bms.com
To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu 
Sent: Tuesday, November 20, 2012 10:03 AM
Subject: [Histonet] Question on frozen section IHC and tissue adhesion

Hi all,
I am performing IHC on tissue that was formalin fixed for 48 hours and then 
embedded in OCT.  I have been using Gold Plus slides from Thermo Fisher.    I 
am staining rat spleen and mandibular lymph nodes.  With a peroxidase method, 
the tissue completely digests off the slides as soon as I apply the peroxidase 
block (looks like pouring hydrogen peroxide on a cut - just turns white and 
bubbles off of the slide).  I have tried various concentrations of peroxidase 
block with no luck.  With the alkaline phos method, the tissue is staying 
adhered to the slide, but I am getting significant 'tissue lifting' which makes 
the pathologists interpretation very difficult.  Is there anyone out there with 
experience in doing IHC on formalin fixed frozen sections that has experienced 
similar 'tissue lifting' problems?  I have ran this antibody on this tissue 
with immunofluorescence successfully - that may be the only choice we have for 
this particular tissue. 
 Would really like to be able to perform this staining with either an HRP or AP 
method for light microscopy if possible.

Christina Thurby
Bristol Myers Squibb
812-307-2093





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delete the message and any attachments. Any disclosure, reproduction, 
distribution or other use of this message or any attachments by an individual 
or entity other than the intended recipient is prohibited.

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[Histonet] question about amplification kit from Ventana

[Gudrun Lang]

Hi!


Can anyone tell me, if the amplification kit of Ventana also works on IgM
antibodies. 

The specification sheet isn't very clear and I must admit, that I just don't
know, if the fc of the IgG and IgM of one species is the same.

In other words, if the anti-IgG in the amplifier also binds to IgM.

 

Thanks in advance

Gudrun

 

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