Can FRAP Be Performed in Water with Small Molecules?

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The discussion centers on the challenges of using Fluorescence Recovery After Photobleaching (FRAP) to measure the diffusion coefficient of small molecules in water. The original poster expresses concerns about the applicability of FRAP in solution, noting a lack of literature on this topic and highlighting difficulties in data evaluation due to the absence of discrete boundaries typical in cellular environments. They mention that the rapid diffusion of small molecules complicates the process, suggesting that techniques like fluorescence correlation spectroscopy may be more appropriate. A reference to a relevant paper on FRAP in cell-free conditions is provided, and the poster acknowledges the potential to derive necessary equations from first principles for their analysis.
Lindsayyyy
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Hello everyone,

I have the following question: I'm doing FRAP in water with a quite small molecule in order to get the diffusion coefficient. I'm quite new to FRAP but I have a feeling that this doesn't work very well. I couldn't find any papers or publications in general about someone doing FRAP in a solution. FRAP seems more like a technique for biological cells. Can anyone confirm this or tell me about papers where they perfom FRAP in solution with low viscosity?

thanks for your helpLindsay
edit: also I did some measurements, but it seems like quite the impossible task to do the evaluation of the data, because I need a whole cell ROI and background ROI which isn't possible in a liquid because I have no discrete borders like I would have in a cell.
 
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thanks for your answer.

Basically I can detect a typical FRAP "spectrum" but I don't know if I can evaulate correctly because of the problems mentioned above (no full 'cell' ROI) and also no background is available to measure.
 
thank you very much, I will take a look
 
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