[Histonet] RE: immunoperoxidase background staining

Troutman, Kenneth A kenneth.a.troutman <@t> Vanderbilt.Edu
Thu Dec 17 13:37:18 CST 2009

A couple of suggestions.  First, what kind of fixative are you using?  If you are using a heavy metal fixative, sometimes that can be the reason.  We have had this issue with B-plus fixative (B5 replacement) and I have seen it with some protocols in zinc formalin and non-buffered fixatives.

Second, try backing off of the CC1 or 2.  I use a Biocare's Background Terminator in an option container to help with my most troublesome antibodies.  You can also use the green diluent they provide you with as a blocking reagent.  I found it works on most things.  If you are having issues with tons of background on ALL your slides, I would send a slide to a place that has established protocols to see if it is the instrument or the tissue.

Hope this helps.

Ashley Troutman BS, HT(ASCP) QIHC
Vanderbilt University Histopathology
1301 Medical Center Drive TVC 4532
Nashville, TN  37232

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