Re: [Histonet] IHC staining intensity.

From:Rene J Buesa

Hi Louise:
  Quantifying IHC staining intensity as an evaluation of the overall reaction
  and epitope abundance has been used in several procedures and 
  consistency on several qualitative aspects of the sections have been identified 
  as the limiting parameter in several procedures.
  There are several papers published in the Journal of Histotechnology that deal
  with this aspect (15: 123-126, 1992; 18: 119-125,1995; 19:23-25,1996; 
  This year I have also published a paper in the JOH [Quantifying Quality: a
  review and scale proposal; 28: 89-97, 2005) that, as the title indicates, deals
  with this subject.
  If you don't have access to my paper, you can e-mail me your FAX number
  and I can send you a copy.
  You are not "logic fuzzy" because there will be cases of cells that have lost
  their "integrity" due to sectioning. The approach would be to define which cells
  or parts of the cells are going to be measured, and only use them.
  Rene J.  
louise renton  wrote:
  Dear all,

I am currently in discussion with someone doing research who is using
a semi quantitative method of reporting stainig using INTENSITY of
cytoplasmic staining as a means of quantifying results. Staining for
a specific antibody is reported as ranging from zero to 3+. What I
would like to know is whether IHC intensity is influenced by the
thickness of the section. What I mean is, if staining is seen in a
whole cell, or one that has been bisected - will there not be a
variation in that one will appear darker when compared to the other?
Or is this variation minimal? Is my logic fuzzy?

I appreciate any input on this matter

best regards

Louise Renton
Bone Research Unit
University of the Witwatersrand
South Africa
"....onwards through the fog!"

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