[Histonet] Blank spots on IHC tissues - bubbles on Bond

[Morken, Timothy]

Joanna, thanks! You are the first who said they had seen it before. I have a Leica tech coming in tomorrow to check out the syringes and lines. That was my thought too.

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

From: Bartczak, Joanna <Joanna.Bartczak at uhn.ca>
Sent: Thursday, August 13, 2020 10:10 AM
To: Paula Keene Pierce <paula at excaliburpathology.com>; Morken, Timothy <Timothy.Morken at ucsf.edu>; Histonet <histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Blank spots on IHC tissues - bubbles on Bond

I have seen this phenomenon years ago on the Bond III when I worked at a different laboratory.  If I remember correctly, Leica had to replace the syringes and lines as air bubbles were being introduced when aspirating reagents.
I would definitely contact Leica technical service and let them investigate.  Good luck.

Sincerely,
Joanna


Joanna Bartczak

Charge Technologist - Immunopathology
University Health Network - Toronto General Site

200 Elizabeth Street

Toronto, Ontario M5G 2C4

Phone:  (416) 340-4800 ext.6004

Joanna.Bartczak at uhn.ca<mailto:Joanna.Bartczak at uhn.ca>



________________________________
From: Paula Keene Pierce <paula at excaliburpathology.com<mailto:paula at excaliburpathology.com>>
Sent: Wednesday, August 12, 2020 6:42 PM
To: Morken, Timothy <Timothy.Morken at ucsf.edu<mailto:Timothy.Morken at ucsf.edu>>; Histonet <histonet at lists.utsouthwestern.edu<mailto:histonet at lists.utsouthwestern.edu>>
Subject: Re: [Histonet] Blank spots on IHC tissues - bubbles on Bond

I have seen this when using Sequenza cover plates.
My thoughts were that it is dissolved gases in the washes. As I noticed that when filling a container with my tap water and leaving it a while, bubbles form on the sides.
Do you brush away bubbles in your water bath even when using distilled water too?
Try not using freshly made, stirred, agitated, etc. solutions and adding a surfactant.
Extreme solution: place solutions in a vacuum.


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On Wednesday, August 12, 2020, 5:25 PM, Morken, Timothy via Histonet <histonet at lists.utsouthwestern.edu<mailto:histonet at lists.utsouthwestern.edu>> wrote:

Has anyone solved a problem on the Leica Bond stainer of bubbles over tissue sections that cause unstained (ie, clear) areas where bubbles have formed?

We've been trying things like extra buffer wash before reagent, new slide trays (tech rep suggestion). It is seen on single- antibody slides and very noticeable on dual - stain slides on which the first stain is there but the second stain shows numerous bubbles that do prevent the second stain from areas of the tissue.  We can see the bubble artifact on the slide away from tissue sections as well.

Any clues to solve this will be appreciated!
Tim Morken
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center

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