FRAP in Water: Can It Be Done?

[Lindsayyyy]

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.

 

[Ygggdrasil]

Yeah, I don't think this will work as the diffusion is probably going to be too fast. Maybe something like fluorescence correlation spectroscopy would be better suited.

 

[Lindsayyyy]

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.

 

[Ygggdrasil]

Here's a paper I found from a quick google search that seems to do FRAP in cell-free conditions: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1300501/

In any case, it should be possible to derive the appropriate equations to use from first principles.

 

[Lindsayyyy]

thank you very much, I will take a look