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

[Paula Keene Pierce]

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. 


Sent from Smallbiz Yahoo Mail for iPhone


On Wednesday, August 12, 2020, 5:25 PM, Morken, Timothy via Histonet <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

_______________________________________________
Histonet mailing list
Histonet at lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet