Posted by
Gabriel Landini on
URL: http://imagej.273.s1.nabble.com/DAB-quantification-tp5006159p5006166.html
On Friday 17 Jan 2014 16:24:44 Panayiota Ploutarchou wrote:
> Thanks for your answer. I agree that I can't quantify, but doesn't darker
> (and more) DAB staining correlate to more antigen present? I was just
> wondering whether there is some sort of semi-quantitative way to go about
> doing this...
No, that is what "non-stoichiometric" means. It is worse than that because DAB
which is used to visualise the result, scatters light rather than transmitting
it and so Beer-Lambert law (to compute the amount of dye based on the optical
density) does not follow well either (so you have 2 problems).
Search the list archives. This has been discussed lots of times. An
interesting post is this one:
https://list.nih.gov/cgi-bin/wa.exe?A2=ind0902&L=IMAGEJ&P=R18412Possible solution: describe what you find with IHC (which cells appear stained
or how many appear stained at a particular antibody dilution) but do not be
tempted to put a numeric quantity to the *intensity* of the stain. Explain why
quantification of DAB intensity is not a good idea and try to find a different
way to quantify what you are after.
That being said, IHC is very useful and has helped immensely in pathology with
tumour typing and clarifying difficult diagnoses, etc. but putting numbers to
the intensity of the stain is stretching the technique too far.
Hope this helps and avoids future headaches.
Gabriel
--
ImageJ mailing list:
http://imagej.nih.gov/ij/list.html