Re: [Histonet] Immunofluorescence problem-Non-specific binding appears in control slide
You may be running into the problem of Falck-Hillarp reactive cells, more
commonly referred to as "SIF" cells. SIF stands for Small Intensely
Fluorescent cells. This technique was quite popular back in the 1960's.
In brief, any cell that contains epinephrine, nor-epi, or Dopamine will
become very highly fluorescent after exposure to formalin. In the actual
Falck-Hillarp technique, the exposure is to gaseous formaldehyde vapor,
but you may be getting some of the same reaction with your procedure.
Generally, Epi and Nor-epi fluoresce with a color that is very similar to
flourescein, while Dopamine has a more orange color.
There is actually a monograph available titled SIF cells. I forget the
exact date, but think it is early '70's. I have a copy on my bookshelf,
but at the moment am a long way from the bookshelf!
Try a Google, Google Scholar or PubMed search for SIF cells - you should
find quite a few publications. It was a very popular technique before
the days of immunostains.
Alton D. Floyd, Ph.D.
23126 South Shore Drive
Edwardsburg, MI 49112
Cell: 574 215-0703
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