Re: [Histonet] IHC troubleshooting

[Greg Dobbin via Histonet]

Nancy,
You used the word “blot” in your reply to Charles, and while I can’t be
certain that you did not mean the same...I would like say in the interest
of clarity, that it is safer to carefully “wick” the water away from under
each section prior to baking.  Blotting (like we might do after a Gram
stain) would likely remove parts or all of the section.
All the best,
Greg
-- 
*Greg Dobbin*
1205 Pleasant Grove Rd
RR#2 York,
PE  C0A 1P0


*Everything in moderation...even moderation itself**!*
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[Histonet] IHC Troubleshooting and Water Baths

[Nancy Schmitt via Histonet]

Hi Charles-

IHC staining - We also use the Leica Bond staining system and had tissue fall 
off in the beginning.  We blot the slides to remove any additional water and 
then place in drying oven for 30 minutes at 60 before placing on the stainer.

Water baths - at the end of each day the water is dumped, the bath is rinsed 
with bleach solution, rinsed with water and dried.

Nancy Schmitt MLT, HT(ASCP)
Pathology Support Services Manager


-Original Message-
From: Charles Riley via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Wednesday, February 26, 2020 9:44 AM
To: Histo List
Subject: [Histonet] IHC troubleshooting

EXTERNAL email from Outside HHC! Do NOT open attachments or click links from 
unknown senders.

Our pathologists are complaining that chunks of the dermis are missing from IHC 
slides yet the entire section is present prior to staining.

Does anyone have any ideas what could cause the tissue to not adhere to the
slides throughout the staining process? We use the Leica Bond stainers.

--

Charles Riley BS  HT, HTL(ASCP)CM

Histopathology Coordinator/ Mohs


Message: 5
Date: Wed, 26 Feb 2020 17:09:14 -0500
From: Charles Riley 
To: Histo List 
Subject: [Histonet] Poll question
Message-ID:

Content-Type: text/plain; charset="UTF-8"

How often does everyone clean their water-bath?

-- 

Charles Riley BS  HT, HTL(ASCP)CM

Histopathology Coordinator/ Mohs



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Re: [Histonet] IHC troubleshooting

[Cartun, Richard via Histonet]

Are you using some form of antigen retrieval?  In addition to what has been 
discussed previously, antigen retrieval using heat (HIER) can cause tissue loss 
especially collagen.

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

-Original Message-
From: Charles Riley via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Wednesday, February 26, 2020 9:44 AM
To: Histo List
Subject: [Histonet] IHC troubleshooting

EXTERNAL email from Outside HHC! Do NOT open attachments or click links from 
unknown senders.

Our pathologists are complaining that chunks of the dermis are missing from IHC 
slides yet the entire section is present prior to staining.

Does anyone have any ideas what could cause the tissue to not adhere to the
slides throughout the staining process? We use the Leica Bond stainers.

--

Charles Riley BS  HT, HTL(ASCP)CM

Histopathology Coordinator/ Mohs
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Re: [Histonet] IHC Troubleshooting

[Greg Dobbin via Histonet]

Hi Charles,
It adhesion problems can arise from several sources, some have already been
mentioned. Here are two that I have experienced:
1) Bad slides. Either a manufacturing defect such that the positive charge
is insufficient or the slides were somehow compromised after opening them
on the bench (perhaps exposed to moisture thus removing the charge).
Sometimes a new lot number be better but sometimes switching brands is the
quickest fix.
2) Ensuring all water is removed from under the section. I find that with
some charged slides (many in fact), water is trapped under the section and
does not drain out on standing and does not drain or evaporate when baking
at 60C. If the slide with trapped water is put on the instrument with
trapped water, the dewax step will lift much of the tissue.
Cheers,
-- 
*Greg Dobbin*
1205 Pleasant Grove Rd
RR#2 York,
PE  C0A 1P0


*Everything in moderation...even moderation itself**!*
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Re: [Histonet] IHC troubleshooting

[Abdulbasit Andijany via Histonet]

In my experience;
If the the specimen is under-fixed or under-processed, the section will be 
sloughed off during IHC staining.

I find proper fixation time and drying the slides at room temperature (before 
baking them in oven) optimal.

Andy



Abdulbasit Andijany (Andy)
AMLS, BBMS, MSc, CT (ASCP)CM, HTL (ASCPI)
King Fahad Armed Forces Hospital, Jeddah, Saudi Arabia


On 26 Feb 2020, at 18:35, John Garratt via Histonet 
mailto:histonet@lists.utsouthwestern.edu>> 
wrote:

Do you float and pick up your sections from a water bath of distilled water 
with no adhesive added? If not, doing that will help.

John

On Wed, Feb 26, 2020 at 6:41 AM, Charles Riley via Histonet 
mailto:histonet@lists.utsouthwestern.edu>> 
wrote:

Our pathologists are complaining that chunks of the dermis are missing from
IHC slides yet the entire section is present prior to staining.

Does anyone have any ideas what could cause the tissue to not adhere to the
slides throughout the staining process? We use the Leica Bond stainers.

--

Charles Riley BS HT, HTL(ASCP)CM

Histopathology Coordinator/ Mohs
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Re: [Histonet] IHC troubleshooting

[John Garratt via Histonet]

Do you float and pick up your sections from a water bath of distilled water 
with no adhesive added? If not, doing that will help.

John

On Wed, Feb 26, 2020 at 6:41 AM, Charles Riley via Histonet 
 wrote:

> Our pathologists are complaining that chunks of the dermis are missing from
> IHC slides yet the entire section is present prior to staining.
>
> Does anyone have any ideas what could cause the tissue to not adhere to the
> slides throughout the staining process? We use the Leica Bond stainers.
>
> --
>
> Charles Riley BS HT, HTL(ASCP)CM
>
> Histopathology Coordinator/ Mohs
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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[Histonet] IHC troubleshooting

[Charles Riley via Histonet]

Our pathologists are complaining that chunks of the dermis are missing from
IHC slides yet the entire section is present prior to staining.

Does anyone have any ideas what could cause the tissue to not adhere to the
slides throughout the staining process? We use the Leica Bond stainers.

-- 

Charles Riley BS  HT, HTL(ASCP)CM

Histopathology Coordinator/ Mohs
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