Re: [Histonet] Von Kossa staining

[Debra Siena via Histonet]

Hi Charles,

I can't answer your question about UV light as I've never tried it except with 
incandescent lights.  However, I do know that you could use natural sunlight as 
well to develop the silver color change.

I hope that helps,
Thanks






Debbie Siena, HT(ASCP)QIHC
Director of Scientific Affairs

Mobile:817-994-9407
Dallas, TX | Baltimore, MD | Mt. Vernon, WA




-Original Message-
From: Charles Riley via Histonet 
Sent: Wednesday, July 26, 2023 1:23 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Von Kossa staining


*** Externally sourced email message ***


Can anyone out there who performs Von Kossa staining provide me with any 
guidelines or suggestions for the light source to use for the Silver nitrate 
activation?

Is a standard handheld black light strong enough or does it need to be a UV 
sanitizing strength light if using UV versus incandescent bulb exposure?
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Re: [Histonet] Alcian Green Dye for Attwood stain

[Debra Siena via Histonet]

Hi John,

Thank you very much. I was hoping that you would also weigh in on this.

Appreciate all your wisdom and willingness to share with others.

Have a great holiday weekend!
Debbie
Get Outlook for iOS<https://aka.ms/o0ukef>

From: John Kiernan via Histonet 
Sent: Thursday, April 6, 2023 12:22 AM
To: 'histonet@lists.utsouthwestern.edu' 
Subject: Re: [Histonet] Alcian Green Dye for Attwood stain


*** Externally sourced email message ***


Alcian green was a mixture of two cationic "Ingrain" dyes: one of ICI's alcian 
blues (a copper phthalocyanine) and their alcian yellow (CI 12840, a monoazo 
dye). ICI stopped making these dyes, which were a commercial flop, in the early 
1970s. It's extremely unlikely that anything now sold as "alcian green" will be 
the genuine article. The original "Alcian blue" dyes were unstable even as 
powders.

The staining procedure published by HD Attwood in J. Path. Bact. 76:211 (1958) 
has to be seen as an oddity from the days when carbohydrate histochemistry was 
still in its infancy. Any textbook published since the 1970s will provide very 
simple and rational staining techniques for showing cartilage matrix 
(basophilic) and keratin (acidophilic) in contrasting colours.

John Kiernan
London, Canada
= = =

To: 'histonet@lists.utsouthwestern.edu' 

Subject: [Histonet] Alcian Green Dye for Attwood stain

Hi All,

I could use some assistance.  I'm gotten a request to do an Attwood stain for 
squames and cartilage and I'm having a difficult time finding the Alcian Green 
2GX dye powder or even a kit if one exists.  Another question is the protocol 
that I've seen from Histonet archives states that you dilute the phloxine in 
cellosolve.  I would just like to ask if anyone has another version of the 
protocol with any updates to the protocol?

Thanks in advance.

Debbie Siena
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From: Debra Siena via Histonet 
Sent: April 5, 2023 2:47 PM
To: 'histonet@lists.utsouthwestern.edu' 
Subject: [Histonet] Alcian Green Dye for Attwood stain

Hi All,

I could use some assistance.  I'm gotten a request to do an Attwood stain for 
squames and cartilage and I'm having a difficult time finding the Alcian Green 
2GX dye powder or even a kit if one exists.  Another question is the protocol 
that I've seen from Histonet archives states that you dilute the phloxine in 
cellosolve.  I would just like to ask if anyone has another version of the 
protocol with any updates to the protocol?

Thanks in advance.

Debbie Siena





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[Histonet] Alcian Green Dye for Attwood stain

[Debra Siena via Histonet]

Hi All,

I could use some assistance.  I'm gotten a request to do an Attwood stain for 
squames and cartilage and I'm having a difficult time finding the Alcian Green 
2GX dye powder or even a kit if one exists.  Another question is the protocol 
that I've seen from Histonet archives states that you dilute the phloxine in 
cellosolve.  I would just like to ask if anyone has another version of the 
protocol with any updates to the protocol?

Thanks in advance.

Debbie Siena





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Re: [Histonet] Breast tissue with calcs

[Debra Siena via Histonet]

Hi Teri,

We have some wonderful new slides they are called MComm slides.   They have 
been well received by our customers, would it be possible to send a box to try 
out?  


 Thanks!
Debbie Siena, HT(ASCP)QIHC
StatLab Technical Support Manager 
 
Phone: 469-525-4949 (O) 817-994-9407 (C)


 
    















-Original Message-
From: Lima, Teresa via Histonet  
Sent: Wednesday, October 26, 2022 2:22 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Breast tissue with calcs


*** Externally sourced email message ***


Hello Histonetters,
We are having issues with keeping micro calcifications in breast biopsy 
sections from falling off our plus slides. Any suggestions would be greatly 
appreciated.
Regards,
Teri

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Re: [Histonet] HSV II control

[Debra Siena via Histonet]

StatLab sells HSV 2 control slides -CSH0525P is the product code and there are 
25 per box.
If I can help in any way, please let me know.

Thanks
 
Debbie Siena, HT(ASCP)QIHC
Technical Support Manager 
Phone: 800-442-3573 option 5 for Technical Support
Dallas, TX | Baltimore, MD | Mt. Vernon, WA
Statlab.com
 
    















-Original Message-
From: Wang, Weixi via Histonet  
Sent: Wednesday, October 12, 2022 8:38 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] HSV II control


*** Externally sourced email message ***


Good morning all,
Does anybody have a control block/slide specific for HSV II? Or any vendor 
providing HSV II control?
Any feedback appreciated.
Thank you,
Weixi

Weixi Wang, Ph.D., HTL(ASCP)
Manager | Histology Laboratory

703 Main Street, Paterson, NJ 07503
Phone: 973.754.3541 | Fax: 973.754.3292
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[Histonet] Ghost Slides

[Debra Siena via Histonet]

Hi All,

I have been asked a question about microscope sides that I have never heard 
before.  Has anyone ever heard the term Ghost Slides?   I would appreciate any 
help if you have heard or understand what this may be referring to.  Thanks so 
much!


[statlab logo]
ISO 13485 Certified

Debbie Siena, HT (ASCP) QIHC
Technical Support Manager
[https://www.statlab.com/media/email/misc/onyourteam.png]
Mobile 817-994-9407 - Fax 972-436-1369
Office: 1-800-442-3573, ext 229
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Re: [Histonet] Retic staining

[Debra Siena via Histonet]

Hi all

Also the freshness of the ammonium hydroxide can make a difference in how much 
you have to add to get the proper reactions. Fresher works better and faster. 
Also I use one of the plastic disposable properties to do drop by drop while 
swirling in s small acid cleaned flask or new plastic container.

Debbie Siena

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From: Cassie P. Davis via Histonet 
Sent: Tuesday, April 23, 2019 1:07 PM
To: histonet
Subject: [Histonet] Retic staining


*** Externally sourced email message ***



Hi Charles,


in addition to the drop size clean glassware makes a big difference. I used to 
use new urine cups to ensure the silver was being made in a clean vessel. I 
hope this helps


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



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[Histonet] Derm IHC question

[Debra Siena via Histonet]

Hello fellow Histonetters
I would like to ask you a question about IHC staining and derm cases.  I am 
seeing a peculiar issue going on, where the melanocytes in the middle of the 
tissues are staining pretty well but when you get to the ends of the tissues 
either shaves or ellipses, they are not staining. This is sporadic, not every 
case and there is no consensus as to a common thread between the cases.I 
feel that this may be a fixation issue but was just wondering if anyone had 
ever seen the same phenomena and would be willing to share the theory or even 
better what was the remedy behind this issue.  The fixative is 10% Neutral 
Buffered Formaliln  and the cells in question that are "dropping out" which is 
what the pathologist is describing are melanocytes, especially with Sox-10 and 
Mart 1 antibodies.
Thanks for the assistance, I definitely appreciate it very much.

Best wishes,

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Empowering Anatomic Pathology
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Re: [Histonet] Poor Staining

[Debra Siena via Histonet]

Hi Charles 

Getting three shades of pink depends a kit on your dehydration steps after 
eosin.  Eosin is more soluble in water than alcohol so you have to watch how 
much water is being absorbed from the atmosphere and change/rotate alcohols 
when you see the last absolute turning pink, also if you have diluted alcohol 
after eosin you may wish to cut down or remove. I don't use a diluted alcohol 
at all just straight absolute alcohol. I recommend 3-4 changes of absolute 
alcohol for 1 minute each. 

For nuclear detail that could be caused by over processing and removing the 
bound water, If this is an occasional occurrence but if happening in all 
tissues large and small, It may also be that your hematoxylin and acid rinse 
need some type of adjustment. 

If I can help you offline please let me know.  Thanks

Debbie Siena, HT(ASCP)QIHC



Sent from my iPhone

> On Jun 16, 2016, at 1:20 PM, Charles Riley via Histonet 
>  wrote:
> 
> Can over processing small biopsies lead to poor chromatin staining? Also
> can this cause an issue with the eosin staining ( for example only getting
> to shades of pink instead of three)?
> 
> Please give me any feedback you can
> 
> -- 
> 
> Charles Riley HT(ASCP)CM
> 
> Histopathology Coordinator/ Mohs
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Re: [Histonet] TC labs

[Debra Siena via Histonet]

They do on my lab which is TC only.

Debbie Siena

Sent from my iPhone

On Oct 5, 2015, at 4:34 PM, Cert NDx 
<cert...@gmail.com<mailto:cert...@gmail.com>> wrote:


I understand that to be the case as well. But in the case of TC only 
laboratories where no full report is being rendered I have yet to see any 
evidence that CLIA knocks on their doors.

On Oct 5, 2015 4:13 PM, "Debra Siena" 
<dsi...@statlab.com<mailto:dsi...@statlab.com>> wrote:
If the lab performs gross examination in any way, the lab must be considered 
high complexity which puts the lab under the umbrella of CLIA and must be 
inspected every two years.

Sent from my iPhone

> On Oct 5, 2015, at 3:54 PM, Andy B via Histonet 
> <histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>> 
> wrote:
>
> Thank you. That is exactly what I would have given as an answer, but I have
> yet to see any CLIA inspector or CLIA Operations manual that mentions any
> of that, other than the vague statements about the Lab Director ultimately
> being responsible. CLIA is not consistent with its rules and regulations
> and interpretations. I would love for someone to prove me wrong!
>
> When is the last time someone actually witnessed CLIA verifying a TC lab
> other than maybe being shown some QC logs, etc? I have yet to see any
> Testing Personnel verifications from CLIA for TC labs. It is hypocritical
> for CLIA not to inspect TC- only operations. If there is a standard form or
> regulation for this, please show me. I have never seen or heard of a PC
> only lab director being asked for verification of testing personnel
> qualifications from a TC lab.
>
> Thanks in advance for everyone's input.
>
>> On Mon, Oct 5, 2015 at 3:42 PM, Andy B 
>> <cert...@gmail.com<mailto:cert...@gmail.com>> wrote:
>>
>> Thank you. That is exactly what I would have given as an answer, but I
>> have yet to see any CLIA inspector or CLIA Operations manual that mentions
>> any of that, other than the vague statements about the Lab Director
>> ultimately being responsible. CLIA is not consistent with its rules and
>> regulations and interpretations. I would love for someone to prove me wrong!
>>
>> When is the last time someone actually witnessed CLIA verifying a TC lab
>> other than maybe being shown some QC logs, etc? I have yet to see any
>> Testing Personnel verifications from CLIA for TC labs. It is hypocritical
>> for CLIA not to inspect TC- only operations. If there is a standard form or
>> regulation for this, please show me. I have never seen or heard of a PC
>> only lab director being asked for verification of testing personnel
>> qualifications from a TC lab.
>>
>> Thanks in advance for everyone's input.
>>
>>
>>
>> On Mon, Oct 5, 2015 at 3:28 PM, Morken, Timothy 
>> <timothy.mor...@ucsf.edu<mailto:timothy.mor...@ucsf.edu>>
>> wrote:
>>
>>> What I believe happens is that the referring lab must do an
>>> audit/inspection of the technical-only lab to be sure they are CLIA
>>> compliant. For instance, check validation procedures for stains, equipment,
>>> quality control. It's true CLIA itself does not inspect the Tech-only lab,
>>> but the referring lab becomes the "technical supervisor" and must have
>>> documentation to show they have inspected it and it is compliant. That is
>>> how it works for vendors using other labs to do work for them under FDA
>>> regulations.
>>>
>>>
>>> Tim Morken
>>> Pathology Site Manager, Parnassus
>>> Supervisor, Electron Microscopy/Neuromuscular Special Studies
>>> Department of Pathology
>>> UC San Francisco Medical Center
>>>
>>>
>>>
>>> -Original Message-
>>> From: Andy B via Histonet 
>>> [mailto:histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>]
>>> Sent: Monday, October 05, 2015 12:05 PM
>>> To: 
>>> histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>
>>> Subject: [Histonet] TC labs
>>>
>>> Does CLIA inspect labs that only perform Technical Components on tissue?
>>>
>>> I know that CAP does so as part of its due diligence, however years ago a
>>> CLIA inspector told me that CLIA doesnt inspect TC-only operations because
>>> there is no report (PC) being rendered. How does this jive with grossing
>>> and the new High Complexity Testing (HCT) paradigm?
>>>
>>> In other words, what is to prevent a pathologist (or non-pathologist)
&

Re: [Histonet] TC labs

[Debra Siena via Histonet]

If the lab performs gross examination in any way, the lab must be considered 
high complexity which puts the lab under the umbrella of CLIA and must be 
inspected every two years. 

Sent from my iPhone

> On Oct 5, 2015, at 3:54 PM, Andy B via Histonet 
>  wrote:
> 
> Thank you. That is exactly what I would have given as an answer, but I have
> yet to see any CLIA inspector or CLIA Operations manual that mentions any
> of that, other than the vague statements about the Lab Director ultimately
> being responsible. CLIA is not consistent with its rules and regulations
> and interpretations. I would love for someone to prove me wrong!
> 
> When is the last time someone actually witnessed CLIA verifying a TC lab
> other than maybe being shown some QC logs, etc? I have yet to see any
> Testing Personnel verifications from CLIA for TC labs. It is hypocritical
> for CLIA not to inspect TC- only operations. If there is a standard form or
> regulation for this, please show me. I have never seen or heard of a PC
> only lab director being asked for verification of testing personnel
> qualifications from a TC lab.
> 
> Thanks in advance for everyone's input.
> 
>> On Mon, Oct 5, 2015 at 3:42 PM, Andy B  wrote:
>> 
>> Thank you. That is exactly what I would have given as an answer, but I
>> have yet to see any CLIA inspector or CLIA Operations manual that mentions
>> any of that, other than the vague statements about the Lab Director
>> ultimately being responsible. CLIA is not consistent with its rules and
>> regulations and interpretations. I would love for someone to prove me wrong!
>> 
>> When is the last time someone actually witnessed CLIA verifying a TC lab
>> other than maybe being shown some QC logs, etc? I have yet to see any
>> Testing Personnel verifications from CLIA for TC labs. It is hypocritical
>> for CLIA not to inspect TC- only operations. If there is a standard form or
>> regulation for this, please show me. I have never seen or heard of a PC
>> only lab director being asked for verification of testing personnel
>> qualifications from a TC lab.
>> 
>> Thanks in advance for everyone's input.
>> 
>> 
>> 
>> On Mon, Oct 5, 2015 at 3:28 PM, Morken, Timothy 
>> wrote:
>> 
>>> What I believe happens is that the referring lab must do an
>>> audit/inspection of the technical-only lab to be sure they are CLIA
>>> compliant. For instance, check validation procedures for stains, equipment,
>>> quality control. It's true CLIA itself does not inspect the Tech-only lab,
>>> but the referring lab becomes the "technical supervisor" and must have
>>> documentation to show they have inspected it and it is compliant. That is
>>> how it works for vendors using other labs to do work for them under FDA
>>> regulations.
>>> 
>>> 
>>> Tim Morken
>>> Pathology Site Manager, Parnassus
>>> Supervisor, Electron Microscopy/Neuromuscular Special Studies
>>> Department of Pathology
>>> UC San Francisco Medical Center
>>> 
>>> 
>>> 
>>> -Original Message-
>>> From: Andy B via Histonet [mailto:histonet@lists.utsouthwestern.edu]
>>> Sent: Monday, October 05, 2015 12:05 PM
>>> To: histonet@lists.utsouthwestern.edu
>>> Subject: [Histonet] TC labs
>>> 
>>> Does CLIA inspect labs that only perform Technical Components on tissue?
>>> 
>>> I know that CAP does so as part of its due diligence, however years ago a
>>> CLIA inspector told me that CLIA doesnt inspect TC-only operations because
>>> there is no report (PC) being rendered. How does this jive with grossing
>>> and the new High Complexity Testing (HCT) paradigm?
>>> 
>>> In other words, what is to prevent a pathologist (or non-pathologist)
>>> from owning and operating a TC lab service that is remote from the PC
>>> service location? How does CMS verify that TC labs are correctly and
>>> compliantly performing grossing (HCT) and other TC lab evolutions if they
>>> do not inspect such operations?
>>> 
>>> I know of labs that are TC only and bill CMS for the Technical Components.
>>> Has anyone else ever wondered about this? How is CMS assuring that TC
>>> labs are compliantly staffed and managed if CMS only inspects labs that
>>> issue reports?
>>> 
>>> Does anyone wonder how CMS can get away with hammering some labs and
>>> ignoring others?
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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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[Histonet] RE: recycled alcohol versus reagent grade alcohol

[Debra Siena]

Hi Ryan,

Reagent grade alcohol can be purchased in different percentages such as 50% to 
100%, etc.  It depends on what you purchase.Reagent alcohol is simply a 
blend of three different alcohols, ethanol, methanol, and isopropyl and is 
created using a formula that is stated by the federal government to make it 
poisonous for human consumption.  

Recycled alcohol never comes back as 100%, and does vary from 95% to about 98%. 
  

  Debbie Siena, HT(ASCP)QIHC
StatLab Medical Products
Technical Support Manager
407 Interchange Street | McKinney, TX 75071
t: 800.442.3573 ext. 229 | f: 972.767.3992
dsi...@statlab.com | www.statlab.com


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Roy, Ryan
Sent: Friday, March 27, 2015 7:53 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] recycled alcohol versus reagent grade alcohol

Hello Histonet,

How do other labs test and verify the percent grade of alcohols and what is the 
assumed percent grade of reagent alcohol? We use recycled alcohols but today 
based on not having reagent grade we used 97% tested recycled as our flush.

The reasoning behind this was that Reagent grade varies from 95%-greater than 
100%. Is this true?

Thanks in advance,

Ryan
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[Histonet] RE: sections not sticking to charged slides

[Debra Siena]

Hi Renee,

Hope you don't mind but I have had a lot of experience in this area.  I worked 
for Ventana for 7 1/2 years as technical specialist and now as a Technical 
Support Manager and  I know that getting sections to stick to slides can be a 
bit challenging especially during antigen retrieval.  One of the rules when 
using positively charged slides is to not use any type of adhesive in the water 
bath, such as Sta-On only use distilled water.  The use of such products 
actually works against you when trying to get the sections adhered to slides.  
Also I would say that when doing IHC, something to keep in mind is that 
adhesive properties of the slides vary over time, so age of the slides is a 
factor and also exposure to humidity and heat is another factor which can 
affect the adhesive properties of the slides.  Make sure that you are not 
buying too many slides at any one time and that you go through them in no more 
than 6 months.  Also if cutting controls and storing in a plastic box that can 
mean that the slides sit around for awhile before being used, so don't cut too 
many controls at one time.  I would suggest drying the slides for at least 30 
minutes to 1 hour at 60C and if you are getting trapped water under the 
section, you may need to dry longer to make sure that all the water is removed. 
 

If you would like to sample some of the StatLab slides that we feel are very 
good for IHC, please let myself or Kevin Collins know, we would be happy to 
sample them if you haven't already tried them.  If I can help in any other way, 
please let me know.  



  Debbie Siena, HT(ASCP)QIHC
StatLab Medical Products
Technical Support Manager
407 Interchange Street | McKinney, TX 75071
t: 800.442.3573 ext. 229 | f: 972.767.3992
dsi...@statlab.com | www.statlab.com

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Renee H. Workman
Sent: Tuesday, February 24, 2015 3:33 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] sections not sticking to charged slides


Help, sections not sticking to charged slides.  We use Mercedes Medical charged 
slides.  Lately have been using sta-on but still have occasional problems 
especially during antigen retrieval.  I need any suggestions.

Renee H. Workman
Histology Supervisor
Virginia Urology
9105 Stony Point Drive
Richmond, VA  23235
W: 804-527-1316 | F: 804-270-0917
rhwork...@uro.com | www.uro.com





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[Histonet] RE: Creutzfeldt-Jakob Disease

[Debra Siena]

Hi Allison,

I would suggest going to the CDC website and pulling from there, they should 
have the latest recommendations.  thanks

Debbie Siena
800.442.3573 ext. 229 | www.statlab.com

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Scott, Allison D
Sent: Tuesday, February 24, 2015 9:45 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Creutzfeldt-Jakob Disease

Hello to all in histoland.  Does anyone have a procedure for handling 
creutzfeldt-jakob disease.  Any help will be appreciated.

Allison Scott HT(ASCP)
Histology Supervisor
LBJ Hospital
713-566-5287(Lab)
713-566-2148(Office)

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[Histonet] best practices for HIER in electric pressure cookers

[Debra Siena]

Hi Histonetters,

I am doing some research into HIER in electric pressure cookers, are there any 
known CAP/CLIA regs about the cooker itself?  Also what is the consensus on 
best practices for using electric pressure cooker these are my questions:

should the amount of slides be standardized per run?
should the amount of water each run be standardized
how about the amount of containers each run?
Temperature strips required each run?
How about releasing the steam early vs letting pressure subside by itself prior 
to opening?
Should HIER times be optimized as well as different pH solutions, for example 
should I test at different time intervals per retrieval solution for best 
results?

Sorry for all the questions but it has been awhile since I have done manual 
retrieval so just want to find out what the guru's are saying.  Thanks in 
advance, if you wish to reply off line that is good and I will publish the 
final results to the histonet.  Thanks again

Debbie Siena
800.442.3573 ext. 229 | www.statlab.comhttp://www.statlab.com/
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Re: [Histonet] RDO

[Debra Siena]

At NSH Mark Ostermeier told us at the awards banquet that it stood for the 
names of his parents, Donald and Rosemary Ostermeier who were the inventors of 
the product that was originally designed to clean pipes. It was interesting to 
learn that bit of trivia.  

Sent from my iPhone

 On Oct 15, 2014, at 6:10 PM, Jennifer MacDonald jmacdon...@mtsac.edu 
 wrote:
 
 Does anyone know what the O signifies in RDO?  Optimal?
 
 Thanks,
 Jennifer
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[Histonet] Eosin leaching

[Debra Siena]

Hi Histonet,


Eosin bleaching is usually due to the alcohols that are used after the eosin, 
if you are in a humid area, the alcohols can actually absorb water from the 
humidity in the air. This may mean a change of how often you rotate or change 
the alcohols, possibly more often than you did during the winter months when 
usually less humid.  Also if you use a graded alcohol after the eosin, maybe 
worth looking into going straight to 100%, I usually always recommend at least 
3 changes 100% alcohol for 1 minute and 3 changes of xylene for 1 minute.  If 
the last alcohol in your staining line is pink, it means that there is eosin 
and that means that there is water.  Eosin loves water and loves to come out of 
solution into the one containing the water.  I have even seen pink xylene in 
some staining lines due to water in xylene, however, not enough that it has 
turned cloudy yet but still have a bit of water.  Xylene will tolerate about 
.5% water before becoming too milky or cloudy to use.  Also when you rotate 
your solutions make sure that you dry the containers well before  refilling and 
by graduating the levels of the alcohols and xylenes to make sure that your 
last container completely covers the slides. Lastly, adding a 95% alcohol 
before your eosin, if you don't have one may help to keep excess water out of 
your eosin which will cause the eosin to break down faster, again eosin loves 
water.  Just spouting off the top of my head but hope something helps out.

Debbie Siena
800.442.3573 ext. 229 | www.statlab.comhttp://www.statlab.com/

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[Histonet] Toluidine Blue stain for Mohs

[Debra Siena]

Hi All,

I am in need of a Toluidine Blue stain for Moh's Lab, would anyone be willing 
to share with me?  It is for a friend.  In advance, thank you very much.

Debbie Siena
800.442.3573 ext. 229 | www.statlab.comhttp://www.statlab.com/

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RE: [Histonet] Negative controls

[Debra Siena]

I just wanted to clarify there are two types of negative controls, one is a 
negative reagent control and one is a tissue that is known negative.  A lot of 
time, we tend to focus on the negative reagent control and forget about the 
negative tissue control.  I know that a lot of smart folks out there on 
histonet will be able to give you some direction as far as how to write a 
policy to cover both but just wanted to clarify that the two types of negative 
controls are different and the negative reagent control may be abandoned if you 
are running a polymer detection kit but the known negative tissue controls are 
still required.  Thanks

  Debbie Siena, HT(ASCP)QIHC
StatLab Medical Products
Technical Support Manager
407 Interchange Street | McKinney, TX 75071
t: 800.442.3573 ext. 229 | f: 972.436.1369
dsi...@statlab.com | www.statlab.com


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Beth Brinegar
Sent: Monday, April 28, 2014 4:56 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Negative controls

Hello fellow histonetters,

What is are other labs doing to satisfy the ANP.22570 QC - Antibodies 
Appropriate negative controls are used.

I apologize, I know this question has been asked before.  I'm trying to satisfy 
these requirements in one procedure.

Thank you all for your assistance!!

Beth Brinegar HTL(ASCP)
Anatomic Pathology Supervisor
Mercy Medical Center
Cedar Rapids, IA 52403
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[Histonet] Retinal detachment

[Debra Siena]

Hi All,

I have a friend that needs help to prevent retinal detachment in mouse eyes.  
Her question is below.  Thanks for any help that you can provide.

We have tried Davidson's fixative before, it is ok. What we are trying to do 
is eliminate the retinal detachment, wherever it comes from (either the 
harvesting technique, fixative, or processing). Will you ask how long do we fix 
mouse and rat eyes in each of the fixatives (alcoholic formalin, zinc formalin 
and Davidson's). After fixation, we transfer to 70% ethanol, and it starts out 
on the processor in 70% ethanol for about a 7 hour processing program. Get back 
to me as soon as you can.  Thank you very much.


Debbie Siena
800.442.3573 ext. 229 | www.statlab.com

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rooster
Sent: Thursday, April 24, 2014 2:08 PM
To: Britton, Josette C
Cc: histonet@lists.utsouthwestern.edu; Duddey, Aimee
Subject: Re: [Histonet] I need opinions on Leica instruments

Love them also!

Chris

On Apr 24, 2014, at 9:11 AM, Britton, Josette C jcbrit...@cheshire-med.com 
wrote:

 I love all of their instruments! Josie Britton, HT (ASCP) QIHC
 
 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of 
 Duddey, Aimee
 Sent: Thursday, April 24, 2014 7:29 AM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] I need opinions on Leica instruments
 
 I am considering the Peloris II tissue processor and the Bond IHC 
 stainer for our lab.  We are a 400 bed regional hospital with a 
 growing outreach business.  We have had Thermo Excelsior processor for 
 years and have never really been happy with it.  Of course our VIP is 
 as reliable and consistent as they come.  We currently have a full 
 Ventana line for IHC (2 ultras, 2 XTs, and BMK special stainer).  I 
 would appreciate any feedback about Leica's line of products.  Please 
 include experiences good and bad.
 Thank you in advance.
 
 Aimee M. Duddey, MLT(ASCP)
 Assistant Director of Laboratory - Pathology FirstHealth Moore 
 Regional Hospital Pinehurst, NC 28374
 (910) 715-5286
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[Histonet] RE: tea bags

[Debra Siena]

StatLab Medical Products sells the paper tea bags, you can contact them at 
800-442-3573.  Thank you

  Debbie Siena, HT(ASCP)QIHC
StatLab Medical Products
Technical Support Manager
407 Interchange Street | McKinney, TX 75071
t: 800.442.3573 ext. 229 | f: 972.436.1369
dsi...@statlab.com | www.statlab.com

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mike Pence
Sent: Friday, December 20, 2013 9:03 AM
To: histonet-boun...@lists.utsouthwestern.edu; histonet@lists.utsouthwestern.edu
Subject: [Histonet] tea bags

Hey all,
Where is everyone getting their tea bags for filtering endo specimens?  I 
cannot find the company that we were using for our bags. They are paper and 
measure 2 ½ x 2 ½ and are sealed on three sides. They come in a strip of bags 
all attached to each other on one side and about 100 to a ziplock bag.

Thanks in advance for the info,
Mike Pence
GRHS
AP Supervisor
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RE: [Histonet] Mixing Paraffin Brands

[Debra Siena]

Hi Lucie,

Paraplast Plus has DMSO and Paraplast Xtra doesn't.  It may be best to 
infiltrate with the Plus which is softer and then embed with the Paraplast Xtra 
which is a bit harder.  I don't usually recommend mixing them especially if you 
only have a few bags.  Thanks

Debbie Siena
800.442.3573 ext. 229 | www.statlab.com

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lucie Guernsey
Sent: Wednesday, June 19, 2013 1:01 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Mixing Paraffin Brands

Hi,

Does anyone know if there's a reason why one shouldn't mix different brands of 
paraffin? We normally use Fisherbrand's Paraplast Plus (melting point 56 C). We 
inherited about 8 bags of McCormick Paraplast X-tra (melting point
52 C). Will the different melting points be a problem?

If we were to use the McCormick paraffin, the only place it may mix with the 
Fisherbrand paraffin is in the blocks themselves (as we refill the embedder). 
But I don't want to compromise the quality of our blocks just to not waste the 
free paraffin.

Or, another option could be that we use the McCormick in the processor and the 
Fisherbrand in the embedder. Could that cause issues in the blocks as the 
tissue would be infiltrated with one brand and embedded in another?

Maybe I'm over-thinking this..

Many thanks!
Lucie

Lucie Guernsey
UCSD
Dept. of Pathology
lguern...@ucsd.edu
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Re: [Histonet] IHC on non-charged slides

[Debra Siena]

Michael

There is a procedure where you can transfer the sections to a charges slide 
with a product called mount quick. You can check with newcomer supply they sell 
it. I know of several labs that have used it on these occasions and it works 
well. 

Sent from my iPhone

On May 16, 2013, at 4:50 PM, Dessoye, Michael J 
mjdess...@commonwealthhealth.net wrote:

 I'm hoping Histonet can come through for me!  I have an unusual case that I 
 need to run IHC on.  Antibody is Pan Keratin Cocktail from Cell Marque with 
 recommended protocol on a Benchmark Ultra with i-view detection.  Only 
 problem is, I only have two slides and they are not charged.  The tissues 
 have been air-dried but not baked.  I cannot obtain more slides.
 
 I'm wondering if there's any kind of pre-treatment I could try to try to help 
 the tissue stay on.  We have run slides like this in the past with mixed 
 results.  Most of the time, the tissue washes off.
 
 Any tips or tricks that might help out in this situation?
 
 
 
 Michael J. Dessoye, M.S. | Histology Supervisor | Wilkes-Barre General 
 Hospital | An Affiliate of Commonwealth Health | 
 mjdess...@commonwealthhealth.net | 575 N. River Street | Wilkes Barre, PA 
 18764 | Tel: 570-552-1432 | Fax: 570-552-1526 
 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _
 
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RE: [Histonet] Re: Home based slide interpretation

[Debra Siena]

It is my understanding that whenever the reading of the slides is not performed 
at the testing site that they need to list that on their report and also they 
will need to have a CLIA license for their home or office that is offsite.

Debbie Siena
800.442.3573 ext. 229 | www.statlab.com

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Wednesday, April 17, 2013 8:35 AM
To: John Spaull; jmacdon...@mtsac.edu
Cc: Histonet
Subject: [Histonet] Re: Home based slide interpretation

John:
You are absolutely right! That is also an issue. Its seems that with every bit 
of additional information the case about being illegal for a pathologist to 
read slides from home is building up more and more strong!
Very interesting.
René J.

From: John Spaull john.r.spa...@gsk.com
To: rjbu...@yahoo.com rjbu...@yahoo.com; jmacdon...@mtsac.edu 
jmacdon...@mtsac.edu 
Sent: Wednesday, April 17, 2013 2:44 AM
Subject: Home based slide interpretation



The UK Human Tissues Act (2006) is also relevant I think. To work with human 
tissue premises have to be registered. All tissue is tracked. The Health 
Service may have exemptions but generally tracking requires approved couriers 
etc. Registration of premises does require payment of a fee.
 
One can imagine the headlines if your home working pathologist was transporting 
slides themselves and say left diagnostic samples on the train. Patient 
confidentiality around physical notes would also be something to be considered 
I guess.
 
--
 
Message: 9
Date: Tue, 16 Apr 2013 06:38:26 -0700 (PDT)
From: Rene J Buesa rjbu...@yahoo.com
Subject: Re: [Histonet] Home based slide interpretation
To: Jennifer MacDonald jmacdon...@mtsac.edu,
  histonet@lists.utsouthwestern.edu
  histonet@lists.utsouthwestern.edu
Message-ID:
  1366119506.58355.yahoomail...@web163104.mail.bf1.yahoo.com
Content-Type: text/plain; charset=iso-8859-1
 
The only regulation about reading slides from home refers to 
cytotechnologists because they have a limit of allowed cases/day that has to be 
controlled?at the usual work setting, but no such regulation exists for 
pathologists and I really?do not think that a pathologist is limited to read 
slides from home.
The only possible objection would be a pathologist working in the UK because, 
under the NHS regulations,? pathologists have norms?about the?time per type of 
case and its complexity.? This time/case was designed to assure that the 
pathologist takes enough time for the diagnosis which, according with the 
system design, assures quality of work, and also links work-to-remuneration 
rates. Under those circumstances perhaps the pathologists would have to justify 
how much time they have used (to be paid for) per different cases, and that 
would require a work based control. 
Fortunately such a system does not exist here (yet!).
Ren? J.
 
From: Jennifer MacDonald jmacdon...@mtsac.edu
To: histonet@lists.utsouthwestern.edu
Sent: Monday, April 15, 2013 6:12 PM
Subject: [Histonet] Home based slide interpretation
 
 
Does anyone know of any regulations against a Pathologist reading slides 
from home?? If so would you be able to cite the specific regulation?
Thanks,
Jennifer
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Regards,
 
John.
 
John Spaull
Histology Group, Molecular and Cellular Technologies, 
Platform Technology and Science,
GlaxoSmithKline RD, Stevenage, SG1 2NY, UK
+44 (0)1438 763296 
E-mail john.r.spa...@gsk.com
 
 

This e-mail was sent by GlaxoSmithKline Services Unlimited
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RE: [Histonet] RE: AFB and negative control

[Debra Siena]

Hi All,

According to CLIA Interpretative guidelines, a negative AFB control tissue is 
to be run each day of testing.

§493.1256 Standard: Control procedures. 

(e)(2) Each day of use (unless otherwise specified in this subpart), test 
staining materials for intended reactivity to ensure predictable staining 
characteristics. Control materials for both positive and negative reactivity 
must be included, as appropriate. 

Interpretive Guidelines §493.1256(e)(2)-(e)(3) 
Acid-fast stains must be checked each day of use for positive and negative 
reactivity.



Debbie Siena
800.442.3573 ext. 229 | www.statlab.com


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of joelle weaver
Sent: Thursday, March 14, 2013 3:51 PM
To: Mayer,Toysha N; 'histonet@lists.utsouthwestern.edu'
Subject: RE: [Histonet] RE: AFB and negative control

This practice is listed as a QC measure for issues of cross contamination in 
the ASCP publication Quality Management in Anatomic Pathology, Nakhleh, R. 
M.D. I have never had any issues that were persistant enough to warrant this 
measure myself, but it is one of the suggestions made under the section for use 
of control tissue/slides. 




Joelle Weaver MAOM, HTL (ASCP) QIHC
  From: tnma...@mdanderson.org
 To: histonet@lists.utsouthwestern.edu
 Date: Thu, 14 Mar 2013 17:52:39 +
 Subject: [Histonet] RE: AFB and negative control
 
 While I don't use a negative control for the AFB, I will use distilled water 
 throughout the procedure.  Most of the time the water in the waterbath is 
 distilled as well, to rule out contamination there as well.  Make sure the 
 waterbath has been disinfected.
 
 Toysha N. Mayer, MBA, HT (ASCP)
 Instructor, Education Coordinator
 Program in Histotechnology
 School of Health Professions
 MD Anderson Cancer Center
 (713) 563-3481
 tnma...@mdanderson.org
 
 
 
 
 Date: Thu, 14 Mar 2013 12:42:30 +
 From: Ian R Bernard ibern...@uab.edu
 Subject: [Histonet] AFB and Negative Control
 To: Lee  Peggy Wenk lpw...@sbcglobal.net, Tighe, Sean T
   sti...@ufl.edu,   histonet@lists.utsouthwestern.edu
   histonet@lists.utsouthwestern.edu
 Message-ID:
   d4f4c602b10b9f45b4e9271af6380e16181a1...@uabexmb1.ad.uab.edu
 Content-Type: text/plain; charset=utf-8
 
 The only special stain that I know that requires the use of a negative 
 control is for the AFB. I understand to rule out false positives as the AFB 
 bacteria might exist in tap water. Nevertheless, a good QA practice which we 
 will implement now.  
 
 Other than Carson, does anyone know of a regulatory or accreditation agency 
 is requiring this as well?  Any suggestion on a good control tissue type? 
 Carson recommends uterus.  Also if there is a pick up on the negative slide 
 (link to the tap water) will use of distilled water and a repeat procedure 
 fix this?
 
 Any thoughts from fellow histonetters?
 
 Thanks
 Ian Bernard
 
 
 
 
 
 
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RE: [Histonet] Retrieving section off of an HE stained slide for immunos

[Debra Siena]

Yes, it is called Mount Quick and Newcomer Supply sells it

Debbie Siena

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Travis Troyer
Sent: Monday, January 14, 2013 2:11 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Retrieving section off of an HE stained slide for immunos

I was wondering if anyone has heard of any method of removing HE stained 
sections from a slide and placing them on a charged slide for immunostaining.

Thanks
Travis
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RE: [Histonet] CAP and negative IHC controls

[Debra Siena]

It is my understanding that you still need a negative tissue control but if 
using a polymer detection kit that you may stop doing a negative reagent 
control.  The negative tissue control is a tissue known to not contain the 
antigen in the tissue, it can be an internal control, or a separate control but 
if internal control, then it must be designated as to how it will be used in 
your procedure manual.  Hope that helps.

Debbie Siena
800.442.3573 ext. 229 | www.statlab.com


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of 
jsjurc...@comcast.net
Sent: Friday, December 28, 2012 2:19 PM
To: .
Subject: [Histonet] CAP and negative IHC controls

How is everyone interpreting the CAP rule about using negative controls? Do we 
still need a negative with each patient slide? 
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[Histonet] Formalin and Operating Rooms

[Debra Siena]

Hi All,

I would like to ask if anyone has heard of any new regulations or laws that 
state that the Operating Room can't have formalin available in the room so that 
they can place formalin onto the sample right away?  I was wondering if anyone 
has heard of this, if you could tell me more about where it is coming from so 
that I can access a copy of it.  We have had some inquiries and I have not 
heard of this.  I appreciate any help that you can give and sorry, that I don't 
have more information.  Best wishes.

  Debbie Siena, HT(ASCP)QIHC
StatLab Medical Products
Technical Support Manager
407 Interchange Street | McKinney, TX 75071
t: 800.442.3573 ext. 229 | f: 972.436.1369
dsi...@statlab.com | www.statlab.com


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[Histonet] RE: counterstain for Alcian Blue (ph2.5)

[Debra Siena]

Hi Diana,

I have heard of using Safranin O as a red counterstain and StatLab sells both 
that and Nuclear Fast Red.   Thanks

Debbie Siena
800.442.3573 ext. 229 | www.statlab.com


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Diana McCaig
Sent: Wednesday, August 01, 2012 11:50 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] counterstain for Alcian Blue (ph2.5)

Can you please let me know what options for counterstain there are other than 
Nuclear Fast Red or a supplier who sells the prepared solution

 

Diana

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RE: [Histonet] Batch Controls

[Debra Siena]

After spending 7 1/2 years as a technical specialist with Ventana, I would also 
like to say that putting the control on the bottom of the slide and the patient 
at the top is just added insurance that the patient will receive the bulk of 
the reagents vs. the control which will help ensure against false negatives 
should there be a staining issue.  Just my 2 cents worth.  thanks

  Debbie Siena, HT(ASCP)QIHC
StatLab Medical Products
Technical Support Manager
407 Interchange Street | McKinney, TX 75071
t: 800.442.3573 ext. 229 | f: 972.436.1369
dsi...@statlab.com | www.statlab.com

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Glen Dawson
Sent: Thursday, April 19, 2012 9:20 AM
To: histonet
Subject: RE: [Histonet] Batch Controls



All,
 
I place a positive control on each slide, next to the patient tissue for all of 
the reasons already mentioned, but we are missing the obvious one.  
 
Many of us use some kind of automated immunostainer where there is no 
gaurantee that, because the CD3 in position #4 (batch control) worked, the 
CD3's loaded in positions 6, 9, 13, and 21 also worked.  Perhaps a reagent ran 
out or there was air in a line for part of the process for any one of these 
other CD3's and, because there is no control on the same slide, there may be a 
false negative result reported due to the use of a batch control.
 
For this reason alone, one should think hard about using batch controls.
 
Just My Opinion,
 
Glen Dawson  BS, HT(ASCP)  QIHC
Histology Technical Specialist
Janesville, WI
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Re: [Histonet] RE: New Reagent Lot Verification

[Debra Siena]

I have heard that you should keep the slides for 2 years or longer if your 
state requires it. 
Debbie Siena HT(ASCP)QIHC
Technical Manager | StatLab Medical Products
407 Interchange St. | McKinney, TX 75071
Direct: 972-436-1010  x229 | Fax: 972-436-1369
dsi...@statlab.com | www.statlab.com 


- Original Message -
From: McMahon, Loralee A [mailto:loralee_mcma...@urmc.rochester.edu]
Sent: Thursday, April 12, 2012 09:54 AM
To: Rathborne, Toni trathbo...@somerset-healthcare.com; 
histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: New Reagent Lot Verification

My interpretation is that you have to compare the new lot to the previous.  So 
I save the slides, so I have the previous to compare it to.   But after that 
lot comparison, I don't know if you should save the slides or not.  I'd like to 
know for sure, since I will surely run out of storage space soon. 
I save the paperwork for the required time as well.

Loralee McMahon, HTL (ASCP)
Immunohistochemistry Supervisor
Strong Memorial Hospital
Department of Surgical Pathology
(585) 275-7210

From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rathborne, Toni 
[trathbo...@somerset-healthcare.com]
Sent: Thursday, April 12, 2012 10:50 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] New Reagent Lot Verification

ANP.22760 refers to new lot verification for antibodies and detection. The 
Evidence of Compliance says that there should be Records of verification of 
new reagent lots. What is everyone's interpretation of records? Are you 
saving slides in addition to paper documentation?


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RE: [Histonet] mesocolon lymphnode treatment with aceton

[Debra Siena]

If anyone has access to the article, I would like to get a copy if you don't 
mind.  thanks

Debbie Siena
800.442.3573 ext. 229 | www.statlab.com

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cynthia Robinson
Sent: Thursday, March 08, 2012 7:55 AM
To: gu.l...@gmx.at; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] mesocolon lymphnode treatment with aceton

I would also be interested if anyone is using this method and their results. I 
have printed off the article and shared the information with our supervising 
pathologist. I have found dermal needle rollers on line but wonder if the 
maximum 2 mm length will do the job of perforating the fat. Also would like 
more information on the compression device used. 

Thanks..Cindi

Cindi Robinson HT(ASCP)
Mercy Medical Center
Dunes Medical Laboratories
350 W Anchor Dr
Dakota Dunes SD 57049
phone-712-279-2768
robin...@mercyhealth.com


 Gudrun Lang gu.l...@gmx.at 3/7/2012 11:24 AM 
Dear all!

Does anyone out there use this method for mesocolon clearing with aceton? I 
heard only a few facts about it and would like to hear about personal 
experience.

The mesocolon is first fixed in NBF, then put in pure aceton, then treated with 
a teasing roller and put in a pipe with holes. The tissue is compressed to 
squeeze out the fat to form a kind of sausage.

The sausage is sliced up, the slices are put into cassettes and processed.

I don't know the exact details. If you have the access to this article, you can 
read more (I don't have). 

http://journals.lww.com/ajsp/Abstract/2012/02000/Optimal_Lymph_Node_Harvest_
in_Rectal_Cancer__UICC.5.aspx

or this:

http://www.springerlink.com/content/748617k331l06143/ 

 

thank you

 

Gudrun Lang

 

Biomed. Analytikerin

Histolab, Linz, Austria

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[Histonet] RE: questions

[Debra Siena]

Hi Dorothy,

I have used reagent alcohol for years both with staining and tissue processing. 
 I have never noticed any artifacts or crystals due to use of reagent alcohol.  
One thing that you should know is that all denatured alcohols are not the same 
so make sure that you buy a denatured (reagent) alcohol of good quality.

As far as keeping slides unstained forever, the one thing that you need to take 
into consideration is that if the slide is charged or silane coated that the 
positive charges (adhesiveness) of the slide will change over time and the 
sections may fall off during staining.  Also the antigenicity of some 
antibodies decreases over time so if you were to do IHC staining, your results 
may not be as strong as they would be on a more recently cut slide.

Best wishes,

Debbie Siena HT (ASCP) QIHC
Technical Manager | StatLab Medical Products
407 Interchange St. | McKinney, TX 75071
Direct: 972-436-1010  x229 | Fax: 972-436-1369
dsi...@statlab.com | www.statlab.com 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Webb, Dorothy L
Sent: Wednesday, February 22, 2012 12:18 PM
To: 'histonet@lists.utsouthwestern.edu'
Subject: [Histonet] questions

For those of you who use reagent alcohol, have you ever experienced any 
problems in processing or staining, such as artifacts, crystals forming, etc??

How long are unstained slides usable?  Do any of you pick up extra sections 
from a ribbon if the tissue is minimal ?  We do and have used them at times, 
but a pathologist would like them saved forever in case tissue is needed for 
a molecular test and there is not enough left in the block.

Appreciate ahead of time your responses!

Dorothy Webb, HT (ASCP)
Regions Hospital



  
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RE: [Histonet] AFB control

[Debra Siena]

It is also a requirement if you are doing work in or for the state of New York. 
 

Debbie Siena
800.442.3573 ext. 229 | www.statlab.com


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mike Pence
Sent: Friday, February 10, 2012 2:49 PM
To: Jennifer MacDonald; Rene J Buesa
Cc: histonet@lists.utsouthwestern.edu; Stephanie Hoyle-Thacker; 
histonet-boun...@lists.utsouthwestern.edu
Subject: RE: [Histonet] AFB control

Recommended by who?
Mike

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jennifer 
MacDonald
Sent: Friday, February 10, 2012 2:43 PM
To: Rene J Buesa
Cc: histonet@lists.utsouthwestern.edu; Stephanie Hoyle-Thacker; 
histonet-boun...@lists.utsouthwestern.edu
Subject: Re: [Histonet] AFB control


It is recommended to run a negative AFB control to rule out false 
positives.  Some sources of water have acid fast organisms, not TB, and if 
the water is used in the water baths or for staining you could pick up 
those organisms.  The recommendation is to cut a section of tissue, known 
NOT to contain AFB organisms, and run it through with the positive control 
and patient.  You would pick it up on the same water bath that you used 
for the patient.




Rene J Buesa rjbu...@yahoo.com 
Sent by: histonet-boun...@lists.utsouthwestern.edu
02/10/2012 11:55 AM

To
histonet@lists.utsouthwestern.edu, Stephanie Hoyle-Thacker 
srhthac...@hotmail.com
cc

Subject
Re: [Histonet] AFB control






All I recall about controls in HC procedures (any of them) is to have a 
(+) control. The only one that requires both (-) and (+) is PAS René J.

--- On Fri, 2/10/12, Stephanie Hoyle-Thacker srhthac...@hotmail.com 
wrote:


From: Stephanie Hoyle-Thacker srhthac...@hotmail.com
Subject: [Histonet] AFB control
To: histonet@lists.utsouthwestern.edu
Date: Friday, February 10, 2012, 1:52 PM



Is anyone using a negative control for AFB stains?  If so is there a 
regulation that states negative controls should be run for AFB stains.  I 
seem to recall seeing such a regulation but cannot find it.

Thanks for your help!

Renee Thacker   
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Re: [Histonet] RE: New CAP question

[Debra Siena]

It is my understanding that one should also revalidate when you switch 
antibodies, detection kits or instruments/methodology.   

As far as validating against another source, you should validate against 
someone doing the same methology, same antibody, and same detection kit. 

Thanks

Debbie Siena HT(ASCP)QIHC
Technical Manager | StatLab Medical Products
407 Interchange St. | McKinney, TX 75071
Direct: 972-436-1010  x229 | Fax: 972-436-1369
dsi...@statlab.com | www.statlab.com 


- Original Message -
From: Podawiltz, Thomas [mailto:tpodawi...@lrgh.org]
Sent: Thursday, August 25, 2011 12:04 PM
To: Laurie Colbert laurie.colb...@huntingtonhospital.com; Vickroy, Jim 
vickroy@mhsil.com; Martha Ward mw...@wakehealth.edu; Carol Bryant 
cb...@lexclin.com; histonet@lists.utsouthwestern.edu 
histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] RE: New CAP question

I might be wrong here but I thought that each time that you brought in new 
testing, you had to validate it at that time, then had to keep those records 
for the life of the test, plus a couple of years. 


Tom Podawiltz HT (ASCP)
Histology Section Head/Laboratory Safety Officer. 



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Laurie Colbert
Sent: Thursday, August 25, 2011 12:46 PM
To: Vickroy, Jim; Martha Ward; Carol Bryant; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] RE: New CAP question

I think it may depend on the inspector.  We had something similar happen
in Cytology during inspection.  They had no validation records for their
Thin Prep processing, which they had been doing for years.  They were
required to validate and provide documentation to CAP.
Laurie Colbert

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Vickroy,
Jim
Sent: Thursday, August 25, 2011 9:31 AM
To: Martha Ward; Carol Bryant; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: New CAP question

I hope you're correct.

James Vickroy BS, HT(ASCP)

Surgical  and Autopsy Pathology Technical Supervisor
Memorial Medical Center
217-788-4046

-Original Message-
From: Martha Ward [mailto:mw...@wakehealth.edu] 
Sent: Thursday, August 25, 2011 11:30 AM
To: Carol Bryant; Vickroy, Jim; histonet@lists.utsouthwestern.edu
Subject: RE: New CAP question

We too have been performing ER and PR for at least 15 years, participate
in CAP proficiency testing and, when we switched staining platforms a
few years ago, validated the new antibody we switched to.   I have
interpreted the standard as necessary if you are introducing ER/PR in
your lab.  In my opinion you would not have to go back and revalidate
something you did years ago just to have something to show at inspection
time.  We had our CAP inspection this summer and a similar question
pertains to the HER2 assay, which we have also been doing for many
years, and that is what I told our inspector, which seemed to satisfy
them. 


Martha Ward, MT (ASCP) QIHC
Manager, Molecular Diagnostics Lab
Dept. of Pathology
Wake Forest Baptist Medical Center
Winston-Salem, NC 27157
336-716-2104

  

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Carol
Bryant
Sent: Thursday, August 25, 2011 12:10 PM
To: 'Vickroy, Jim'; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: New CAP question

Please respond to all.  I would like the information also. 
Thank you,
Carol 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Vickroy,
Jim
Sent: Thursday, August 25, 2011 12:01 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] New CAP question

One of the new CAP questions is ANP.22976 ER/PgR validation.

If the laboratory performs immunohistochemistry for estrogen receptor
and/or progesterone receptor as a prognostic/predictive marker on breast
carcinoma, the laboratory has documented appropriate validation for the
assays.  In the note it says should include a minimum of 40 cases and
validation should be performed by comparing the laboratory's results
with another assay that has been appropriately validated.

We have been doing ER/PR's for over ten years.  Originally we compared
our ER/PR testing with the old immunology method that used frozen breast
tissue.   We also compared our ER/PR results with another hospital.
Problem is that this has been over ten years and we do not keep quality
control records that long.   Am I missing something?
I know we use the FDA approved protocol from Ventana on our Ventana
Benchmark XT.
Should we do another validation study using Ventana or another hospital
that is using the FDA approved method?   Anybody understand what CAP is
wanting and how to accomplish this?

James Vickroy BS, HT(ASCP)

Surgical  and Autopsy 

Re: [Histonet] Where Can I Buy Pneumo Control Slides

[Debra Siena]

Statlab sells pneumo controls and they are in human tissues. 

Thanks
Debbie Siena HT(ASCP)QIHC
Technical Manager | StatLab Medical Products
407 Interchange St. | McKinney, TX 75071
Direct: 972-436-1010  x229 | Fax: 972-436-1369
dsi...@statlab.com | www.statlab.com 


- Original Message -
From: Laurie Colbert [mailto:laurie.colb...@huntingtonhospital.com]
Sent: Monday, July 25, 2011 04:21 PM
To: wanda.sm...@hcahealthcare.com wanda.sm...@hcahealthcare.com; 
histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Where Can I Buy Pneumo Control Slides

American Master Tech.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
wanda.sm...@hcahealthcare.com
Sent: Monday, July 25, 2011 1:26 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Where Can I Buy Pneumo Control Slides

Good Afternoon to All,
I am getting pretty desperate to find a source for Pneumo control
slides.  I know they are hard to come by, but my favorite source
Newcomer Supply does not have any and I ordered some from Sigma-Aldrich
and now they are discontinued.  Please Help!
Thanks,
Wanda

WANDA G. SMITH, HTL(ASCP)HT
Pathology Supervisor
TRIDENT MEDICAL CENTER
9330 Medical Plaza Drive
Charleston, SC  29406
843-847-4586
843-847-4296 fax


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RE: [Histonet] Slide Bright Question

[Debra Siena]

Hi Christopher,

There are differences between how Xylene substitutes works and Xylene.  First 
of all, Xylene subs can be a bit slower to deparaffinize, so you may need to 
increase the time or # or stations when deparaffinizing.  Also, Xylene subs 
don't tolerate water so your last alcohol must be 100% or else you will see the 
eosin bleed out of the sections.  The other thing about Xylene substitutes is 
that the mounting media must be compatible with the substitute.   If I can help 
in any other way, please let me know.  thanks

Debbie Siena HT(ASCP)QIHC
Technical Manager | StatLab Medical Products
Direct: 972-436-1010  x229 



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Christopher 
Conlisk
Sent: Friday, June 03, 2011 1:18 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Slide Bright Question

Hello Everyone,
I have worked in labs that use Xylene my entire career and I just
started at a Lab That only uses a Xylene Substitute Slide-Bright. I am
having problems with the HE. After staining and coverslipping (The slides
look fine innitially), Then about 5-10 minutes after coverslipping the Eosin
starts bleeding out all around the tissue. I have asked several of my
Histotech Friends that are old timers and they say that Xylene Substitutes
are awful at deparrifinization and awful at clearing. They told me that the
alcohol isnt getting thoroughly cleared in the Slide Brite and then it is
eventually leeching out after coverslipping??? Is this true and does anyone
have any guidance for this issue? We also run MOHS slides on the same
stainer and I keep all the reagents clean as a whistle. I really hate Xylene
Substitute's

Thanks

C.S. Conlisk HT(ASCP), PBT(ASCP)
Kansas City Skin and Cancer Center
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[Histonet] DMSO in Tissue processor

[Debra Siena]

Hi Everyone,

I was hoping that I could ask a question about the use of paraffins with DMSO 
in tissue processors.  I was wondering if there are any tissue processors that 
recommend or specify that paraffins containing DMSO not be used.  I am not 
aware of any of the older versions but just wondered about the newer microwave 
processors and other newer versions of processors now on the market.  Thanks 
for all your help!

Debbie Siena HT(ASCP)QIHC
Technical Manager | StatLab Medical Products
407 Interchange St. | McKinney, TX 75071
Direct: 972-436-1010  x229 | Fax: 972-436-1369
dsi...@statlab.commailto:maub...@statlab.com | 
www.statlab.comhttp://www.statlab.com/


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RE: [Histonet] clia reg on ventilation

[Debra Siena]

You may want to check out Dick Dapson's book on Laboratory Safety, there is a 
chapter on ventilation there.

Dapson R.W. (2001) Safety in the laboratory, Chapter 6 in Theory and Practice 
of Histological
Technique Ed by Bancroft J.D. Gamble, M Elsevier Science Ltd

Debbie Siena HT(ASCP)QIHC
Technical Manager | StatLab Medical Products
Direct: 972-436-1010  x229 


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Nicole Tatum
Sent: Tuesday, April 05, 2011 9:22 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] clia reg on ventilation

Can anyone help me find some kind of standard for ventilation in a
pathology laboratory. I cant seem to find anything more then adequate
ventilation.  Ok well what definies adequate? Is there a set Clia standard
or AHCA standard for ventilation in Flordia. Thank you for your help.

Nicole


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[Histonet] Diff Quik or Hema Diff on Frozen Sections

[Debra Siena]

Hi All,

If anyone is using the Diff Quik or Hema Diff stains on frozen sections, would 
you mind sharing your protocol, especially in regards to how the FS slides are 
handled after cutting, are they air dried, or not and what fixative if any do 
you use before staining.  Thanks in advance, I appreciate all the helpful 
information that I receive on Histonet.

Debbie Siena HT(ASCP)QIHC
Technical Manager | StatLab Medical Products
407 Interchange St. | McKinney, TX 75071
Direct: 972-436-1010  x229 | Fax: 972-436-1369
dsi...@statlab.commailto:maub...@statlab.com | 
www.statlab.comhttp://www.statlab.com/


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[Histonet] bone marrow aspirations

[Debra Siena]

Hi All,

I would appreciate some insight as to how most labs are treating bone marrow 
smears, in regards to fixation.  I am not embarrassed to say that it has been 
awhile since I worked with them.  With the smears are most labs air drying, 
methanol (alcohol fixation) or spray fixation (pap fixative) or something else? 
 I do thank you for your help.

Debbie Siena HT(ASCP)QIHC
Technical Manager | StatLab Medical Products
407 Interchange St. | McKinney, TX 75071
Direct: 972-436-1010  x229 | Fax: 972-436-1369
dsi...@statlab.commailto:maub...@statlab.com | 
www.statlab.comhttp://www.statlab.com/


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[Histonet] Bone marrow smears

[Debra Siena]

Hi All,

It appears that air drying and then methanol fixation is the method most labs 
are using on bone marrow smears.  Thanks for all your help, I do appreciate all 
the responses.  Best Wishes

Debbie Siena HT(ASCP)QIHC
Technical Manager | StatLab Medical Products
407 Interchange St. | McKinney, TX 75071
Direct: 972-436-1010  x229 | Fax: 972-436-1369
dsi...@statlab.commailto:maub...@statlab.com | 
www.statlab.comhttp://www.statlab.com/


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[Histonet] Diff Quik/Wright stain for stool samples

[Debra Siena]

Hello All,

Does anyone use a Diff Quik protocol for parasites in stool samples and if so 
could you send me a copy of your procedure?  I would greatly appreciate it.  
Thank you all.

Debbie Siena HT(ASCP)QIHC
Technical Manager | StatLab Medical Products
407 Interchange St. | McKinney, TX 75071
Direct: 972-436-1010  x229 | Fax: 972-436-1369
dsi...@statlab.commailto:maub...@statlab.com | 
www.statlab.comhttp://www.statlab.com/


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RE: [Histonet] TTF-1/background staining

[Debra Siena]

Hi Paula,

I am probably reading between the lines here but on your TTF protocol do you 
use a streptavidin biotin detection system and if so do you block for 
endogenous biotin, liver will have endogenous biotin where lung may not have as 
much?  thanks

Debbie Siena HT(ASCP)QIHC
Technical Manager | StatLab Medical Products
Direct: 972-436-1010  x229 


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Paula Lucas
Sent: Thursday, February 03, 2011 2:38 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] TTF-1/background staining

How can I eliminate the background or as you say, non-specific staining?

 

I'm no expert here, I admit, and so I'm asking for your help and
suggestions.  I have searched the Histonet archives, and a lot of what I'm
seeing deals with animal or rodent tissues, and a lot of it was confusing to
me.

 

To give you a little background info:

We use the Lab Vision stainer and the TTF-1 antibody we use is from Cell
Marque.  We were having issues with this marker from Lab Vision, so we
switched to TTF-1 a while ago.  We use the UltraVision LP detection kit from
Lab Vision. It's a polymer driven detection kit. 

 

The antibody is a ready to use, and we have the time set at 30 minutes.  

 

We were getting the background staining on a liver specimen last week, and
we also had this problem today on a lung case.  My doctor is getting
frustrated, and wants me to do something about this, and to repeat the TTF-1
stain on the lung, so if someone can give me some suggestions to try, I'm
ready to try them.

 

Thanks in advance,

Paula

Lab Manager

Bio-Path Medical Group

Fountain Valley, CA

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[Histonet] Marvin Hanna

[Debra Siena]

Hi All,

Sorry to do this on the list but if Marvin Hanna is out there, could you 
contact me off line.  Thanks

Debbie Siena HT(ASCP)QIHC
Technical Manager | StatLab Medical Products
407 Interchange St. | McKinney, TX 75071
Direct: 972-436-1010  x229 | Fax: 972-436-1369
dsi...@statlab.commailto:maub...@statlab.com | 
www.statlab.comhttp://www.statlab.com/


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RE: [Histonet] HE Stain

[Debra Siena]

Hi Toni,

I would love to speak with you about the issues that you are having, could you 
give me a call?   I am traveling tomorrow but will be back in the office on 
Friday.  thanks

Debbie Siena
Technical Manager | StatLab Medical Products
Direct: 972-436-1010  x229
800-442-3573 ext 229


 

 


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rathborne, Toni
Sent: Wednesday, January 19, 2011 12:35 PM
To: Mike Pence; Scott, Allison D; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] HE Stain

We have been having the same problem recently. We have tried extending the 
washes after the hematoxylin, agitation and adding an additional wash. Nothing 
has helped. It is not every slide as you say, but random ones. We are using 
Gill 3 and eosin from Stat Lab. We have changed the stainer and processor a 
number of times since this began.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu]On Behalf Of Mike
Pence
Sent: Wednesday, January 19, 2011 1:10 PM
To: Scott, Allison D; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] HE Stain


More often than I would like to admit when I have seen this type of
problem it has been that there is a solution out of place on the
processor or the stainer. I would start there.
Mike

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Scott,
Allison D
Sent: Wednesday, January 19, 2011 11:59 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] HE Stain


Hello to all in histoland and Happy New Year.  We are having issues with
our HE stain.  The nuclei are staining very blue to purple and the
mucin is staining blue to purple-blue.  It is difficult to see the
nuclear detail.  The mucin is obscuring things.  We have not changed our
process for staining or processing.  The funny thing is that it is only
in the Biopsy cases, and it is every few slides.  The surgical  cases
are all right.  We checked the alcohol and xylene for water, and there
is not any.  My tech changed out the stain and we are staining a new
batch of slides.  If anyone has any idea what is wrong, any help would
be greatly appreciated.  I have gone over our processes and nothing has
changed.  The reagents are the same, the staining times are the same,
and the processing times are the same.  We are using the Shandon Gemini
stainer and VIP processor.

Allison Scott HT(ASCP)
Histology Supervisor
LBJ Hospital
Houston, Texas
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[Histonet] RE: Looking for a product

[Debra Siena]

Hi Kaye,

StatLab Medical Products sells the mini-fad pads and our customer service 
number is 800-442-3573.  Thanks

Debbie Siena HT(ASCP)QIHC
Technical Manager | StatLab Medical Products
Direct: 972-436-1010  x229 


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kaye Ryan
Sent: Tuesday, January 18, 2011 11:20 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Looking for a product

Hi Everyone,

I am looking for a product call mini fan pads.  They are a pad that
will neutralize formalin.  I can't remember who sells them.  Any help
would be appreciated.  Last time I saw them they came like a roll of
paper towels.

 

Thanks in advance,

Kaye Ryan

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