Re: [Histonet] Staining issues

[Tony Henwood (SCHN) via Histonet]

Hi Charles,

Many issues could be in play.
Since the centre of the tissue is causing problems, then I would look at 
fixation; are the spaces around cells in the centre of the block wider than at 
the edges (conversely do the cells look like they are smaller than their 
counterparts at the edge)?

If part of this tissue is lightly fixed, they seem to be more prone to heating 
effects. If the haematoxylin is paler (washed-out?) in the centre then it is 
possible that the slides have been heated above 70oC before H staining. This 
will also affect the IPX's. Maybe the oven is over-heating.

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Principal Scientist, the Children's Hospital at Westmead
Adjunct Fellow, School of Medicine, University of Western Sydney 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 

-Original Message-
From: Charles Riley via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Thursday, 29 December 2016 12:19 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Staining issues

Our pathologists are complaining that the center of tissues are not staining 
properly in both our H's and IHC's. Can anyone provide some thoughts as to 
where this problem could be occurring?


As a separate situation we are experiencing the tissues folding or falling off 
during the staining process. Both this issue and the staining issue are recent 
occurrences and we have not changed our process in any way from previous years.


Does anyone think these problems are related? Why or why not?   I have
racked my brain with all the trouble shooting techniques I can think of so any 
help will greatly be appreciated

-- 

Charles Riley HT(ASCP)CM

Histopathology Coordinator/ Mohs
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[Histonet] FJC controls

[Michele Wich via Histonet]

Does anyone have any good Fluoro Jade C controls that they would be willing to 
sell or possibly trade for some other control blocks?

Thanks,
Michele
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Re: [Histonet] Histonet Digest, Vol 157, Issue 21 the center

[Steve McClain via Histonet]

What paraffin are you using?
I heard a rumor one manufacturer changed formulations recently. Same brand name 
different formula. 

Center problems often  manifest where  insufficient fixation is a primary event 
or where rapid processing meets the limits of solution maintainance and tissue 
size. Rapid processing w inadequate alcohols is a silent offender. Sometimes 
you just replace every solution, just so you know where you a new starting 
point. 

If you suspect a processing problem as a cause, one can often achieve a better 
result by melting and re-embedding in new paraffin. By this I mean Allow to sit 
in the embedding center melted in a new mold, in fresh new paraffin for 
60minutes. Then Change paraffin again and embed and cool. Not sure why it 
works, but cooking for a few more minutes does.improve sectioning in marginal 
tissues.

Pathologists can also be the cause- Tissues which are crushed have aberrant 
staining patterns. 
XS handling prior to fixation can mess up staining. Raw tissues rushed through 
and on the processor. I am guilty of all of them in the past.

Steve A. McClain, MD


> On Dec 28, 2016, at 13:30, "histonet-requ...@lists.utsouthwestern.edu" 
>  wrote:
> 
> Our pathologists are complaining that the center of tissues are not
> staining properly in both our H's and IHC's. Can anyone provide some
> thoughts as to where this problem could be occurring?
> 
> 
> As a separate situation we are experiencing the tissues folding or falling
> off during the staining process. Both this issue and the staining issue are
> recent occurrences and we have not changed our process in any way from
> previous years.

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Re: [Histonet] CAP checklist help

[Walter Benton via Histonet]

Leica sells a temperature verification kit that should meet your needs for CAP 
compliance and documentation. We use it for our Leica Thermobrites used in the 
processing of FISH.

-Original Message-
From: Charles Riley via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Wednesday, December 28, 2016 11:55 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] CAP checklist help

I need help with figuring out how to meet checklist # ANP.23420.   We use
the Leica Bond III and MAX  to do our ISH slides.  How do I test the 
temperature and how would you recommend recording the results?

--

Charles Riley HT(ASCP)CM

Histopathology Coordinator/ Mohs
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Re: [Histonet] CAP checklist help

[Eddie Martin via Histonet]

Hi Charles,

The Leica BOND system displays the temperatures of the individual heating pads 
during the retrieval stage on your computer monitor. You can easily take a 
screenshot of your computer monitor and save it to a flash drive as a digital 
record.
*A heating error would appear if the individual heating pad didn't reach 100 
degrees. 

 The Leica service engineer for the Bond can also provide documentation every 
time they service your IHC analyzer. You can also check your slide staining 
history and find cases that didn't stain well and individually check if the 
result was due to the temperature not reaching 100 degrees on that individual 
heating pad. Hope this helps. 

V/r,
Eddie Martin, HTL, QIHC
IHC Histotechnologist III
Walter Reed National Military Medical Center 


> On Dec 28, 2016, at 11:55 AM, Charles Riley  wrote:
> 
> I need help with figuring out how to meet checklist # ANP.23420.   We use
> the Leica Bond III and MAX  to do our ISH slides.  How do I test the
> temperature and how would you recommend recording the results?
> 
> -- 
> 
> Charles Riley HT(ASCP)CM
> 
> Histopathology Coordinator/ Mohs
> 

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[Histonet] CAP checklist help

[Charles Riley via Histonet]

I need help with figuring out how to meet checklist # ANP.23420.   We use
the Leica Bond III and MAX  to do our ISH slides.  How do I test the
temperature and how would you recommend recording the results?

-- 

Charles Riley HT(ASCP)CM

Histopathology Coordinator/ Mohs
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Re: [Histonet] Staining issues

[Rene J Buesa via Histonet]

Improper staining at the center and falling sections are typical consequences 
of poor fixation/infiltration.If you have changed nothing proceduraly, what 
about somebody "new" grossing and preparing thicker tissue slices?René 

On Wednesday, December 28, 2016 8:34 AM, Charles Riley via Histonet 
 wrote:
 

 Our pathologists are complaining that the center of tissues are not
staining properly in both our H's and IHC's. Can anyone provide some
thoughts as to where this problem could be occurring?


As a separate situation we are experiencing the tissues folding or falling
off during the staining process. Both this issue and the staining issue are
recent occurrences and we have not changed our process in any way from
previous years.


Does anyone think these problems are related? Why or why not?  I have
racked my brain with all the trouble shooting techniques I can think of so
any help will greatly be appreciated

-- 

Charles Riley HT(ASCP)CM

Histopathology Coordinator/ Mohs
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[Histonet] Staining issues

[Charles Riley via Histonet]

Our pathologists are complaining that the center of tissues are not
staining properly in both our H's and IHC's. Can anyone provide some
thoughts as to where this problem could be occurring?


As a separate situation we are experiencing the tissues folding or falling
off during the staining process. Both this issue and the staining issue are
recent occurrences and we have not changed our process in any way from
previous years.


Does anyone think these problems are related? Why or why not?   I have
racked my brain with all the trouble shooting techniques I can think of so
any help will greatly be appreciated

-- 

Charles Riley HT(ASCP)CM

Histopathology Coordinator/ Mohs
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