Hello everyone,
Does anyone know whether collagen-cell binding can somehow alter the fluorescence signal from collagen-FITC conjugate and why? I am working with plastic petri dishes coated with collagen Type I-FITC conjugate and I noticed a change in fluorescence intensity in the presence of adherent epithelial cells.
I initially thought it might be associated with ions released by cells, but this does not seem to be the case. Can some kind of structural change in the collagen affect the fluorescent signal?
Dear Stefano Villa In my own experience FITC fluorescence (long time ago but still) is sensitive to multiple factors. To exclude some of them you might consider the use of things like:
FITC labeled-polyLysine
FITC-Dextrane
Adding unlabeled collagen
You might consider having a look at
Chapter Characterization of helical junction zones in gelatin networ...
especially the ‘Fluorescence measurements’ section for details.In other words, try to understand the system you use when it comes to the FITC fluorescence behavior. Are you measuring what you want and are the changes you see caused by what you expect etc.
Best regards.
Dear Rob Keller ,
thank you for the answer. Somehow the reference that you recommend is not visible in your answer. Can you write it again?
Dear Stefano Villa Have another look. You were to quick ;-) I needed to add the link after I replied.
It should be visible now.
Kind regards, Rob
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