[Histonet] pretreatment for IHC

[Tan, MinHan]

I think that the appropriate thing to do for increased background,
rather than attempt all kinds of methods, is to elicit the step at which
problems are occurring by use of appropriate controls. This will save
you both time and money ultimately.


You can run all these in a single experiment - just make sure that the
slides are cut from the same specimen, which is known to stain positive.


 

There are quite a few causes of increased background, of course - but
this will allow you to find out with the minimum of fuss, and within one
day. 

 

Slide with substrate alone

Slide with substrate + ABC

Slide with substrate + ABC + secondary antibody

Slide with substrate + ABC + primary antibody + secondary antibody 

Slide with substrate + ABC + primary antibody + secondary antibody +
peroxidase quenching

Slide with substrate + ABC + primary antibody + secondary antibody +
blocking reageants

Slide with substrate + ABC + primary antibody + secondary antibody +
peroxidase quench + blocking reageants. (dissimilar species serum)

 

Min-Han Tan

Van Andel Research Institute

 

 

-----Original Message-----
From: Scholz, Stephen J. [mailto:Stephen.J.Scholz <@t> osfhealthcare.org] 
Sent: Friday, December 05, 2003 12:32 PM
To: Histonet <@t> Pathology.swmed.edu
Subject: [Histonet] pretreatment for IHC

 

Hello Histonet, 

I am having a problem with background staining on IHC slides.  This is
only happening in cell block specimens.  Is this common on cell blocks?
Should I do a pretreatment on these?  What pretreatments do others
prefer?  Should I peroxide quench?  Before or after?  What about
blockers?  Do I ramble on and ask to many questions?

Thanks for all your help, 

Stephen J. Scholz HT(ASCP) 
Histology Coordinator 
OSF St. Anthony Medical Center 
Rockford IL 

Phone: 815-395-5410 
Fax: 815-395-5364 
e-mail: sjscholz <@t> osfhealthcare.org 


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