Can FRAP Be Performed in Water with Small Molecules?

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In summary, the conversation is about someone who is new to FRAP (fluorescence recovery after photobleaching) and is trying to measure the diffusion coefficient of a small molecule in water. They are unsure if this technique will work in a solution and are looking for papers or publications where FRAP has been performed in low viscosity solutions. Another person suggests using fluorescence correlation spectroscopy instead. One person has trouble evaluating the data due to the lack of a full cell ROI and background ROI in a liquid. However, a paper was found that uses FRAP in cell-free conditions and it should be possible to derive the appropriate equations from first principles.
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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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  • #3
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.
 
  • #5
thank you very much, I will take a look
 

FAQ: Can FRAP Be Performed in Water with Small Molecules?

Can FRAP be used to measure antioxidant activity in water?

Yes, FRAP (Ferric Reducing Antioxidant Power) is a commonly used method to measure antioxidant activity in various substances, including water.

How does FRAP work to measure antioxidant activity?

FRAP measures the ability of a substance to reduce iron (Fe3+) to its ferrous (Fe2+) form. This reduction is caused by the presence of antioxidants in the substance, which donate electrons to the iron ions, resulting in a color change that can be measured and quantified.

What are the limitations of using FRAP to measure antioxidant activity in water?

FRAP is a simple and inexpensive method, but it has some limitations. It only measures the reducing capacity of antioxidants and does not take into account other properties that may contribute to their overall antioxidant activity. Additionally, the results may vary depending on the type and concentration of antioxidants present in the water.

Is FRAP suitable for testing all types of water?

FRAP can be used to measure antioxidant activity in various types of water, including tap water, bottled water, and natural water sources. However, the presence of certain compounds or contaminants in the water may interfere with the results, so it is important to consider the potential sources of error before conducting the test.

Are there other methods for measuring antioxidant activity in water?

Yes, there are other methods for measuring antioxidant activity in water, such as ORAC (Oxygen Radical Absorbance Capacity), DPPH (2,2-diphenyl-1-picrylhydrazyl), and TEAC (Trolox Equivalent Antioxidant Capacity). Each method has its own advantages and limitations, and the choice of method may depend on the specific research or application needs.

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