[Histonet] RE: Artifact in IHC

Geoff mcauliff <@t> rwjms.rutgers.edu
Mon Sep 22 13:40:53 CDT 2014

Teri makes an excellent point about higher alcohols causing 
precipitation of phosphate salts.


On 9/22/2014 1:17 PM, Teri Johnson wrote:
> Hi Donna,
> Interesting conundrum! A few questions:
>   - Do you see it in tissues that have not undergone any staining, just floated in buffer and coverslipped?
>   - Have you tried wet mounting them after staining and prior to dehydration and clearing to see if the artifact is there?
>   - Have you recently changed glove manufacturers, could this be talc or some other powder from gloves?
>   - Are you dehydrating out of PBS into an alcohol concentration higher than 70%? You could be getting phosphate precipitation. Try using a water rinse prior to alcohol or make sure your first alcohol concentration is 70% or lower.
> Those are the weird things that came to mind. I hope this helps,
> Teri Johnson
> Manager Clinical Trial Testing
> Genoptix, Inc.
> Carlsbad, CA 92008
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732) 235-4583; fax: -4029
mcauliff <@t> rwjms.rutgers.edu

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