Can FRAP Be Performed in Water with Small Molecules?

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Discussion Overview

The discussion centers around the feasibility of performing Fluorescence Recovery After Photobleaching (FRAP) in water with small molecules to determine diffusion coefficients. Participants explore the applicability of FRAP in liquid solutions, particularly in the context of low viscosity environments, and the challenges associated with data evaluation in such scenarios.

Discussion Character

  • Exploratory, Technical explanation, Debate/contested

Main Points Raised

  • One participant expresses uncertainty about the effectiveness of FRAP in liquid solutions, noting a lack of literature on the subject and suggesting that FRAP is primarily used for biological cells.
  • Another participant argues that the diffusion in liquid may be too rapid for FRAP to yield useful results, proposing fluorescence correlation spectroscopy as a potentially better alternative.
  • A participant mentions detecting a typical FRAP spectrum but questions the validity of data evaluation due to the absence of a full cell region of interest (ROI) and background measurements in a liquid medium.
  • One participant shares a reference to a paper that reportedly discusses FRAP in cell-free conditions, suggesting that it may be possible to derive relevant equations from first principles.

Areas of Agreement / Disagreement

Participants do not reach a consensus on the viability of performing FRAP in water with small molecules. Multiple competing views are presented regarding the technique's applicability and the challenges faced in data evaluation.

Contextual Notes

Limitations include the dependence on specific conditions for FRAP to be effective, the need for discrete borders for ROI in liquid solutions, and unresolved questions about the appropriate equations for 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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