FRET analyze

[Pablo Munoz-2]

> Dear members.
>
> I have a big problem. I need to analyze several FRET  images (time lapse videos) before and after of a treatment. If a separate the image from the video are like 10 pictures by CFP, YFP and FRET channel. So is a lot to manage. My interest is measure a ROI in the tip of a neurite during the time. I used a cAMP probe. So i want to see the changes during the time.
>
> Does anybody know or have a macros or a idea to simplify my analyze , Please?
>
> Thanks in advance for all your help
>
> Kind Regards
>
> Pablo Munoz Ll
>

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[Nguyen P.]

Pablo:

Since you are using a known cAMP probe, you can directly compare and plot the FRET intensity values at the neurite tip for each of the images. Alternately you can either look at the fluctuations of YFP or the YFP:CFP ratios for the ROI of interest. For more complicated measurements you can use normalized FRET which you dont necessarily have to use.

I would use imageJ define the roi and obtain fl intensity measurements for them. Usng the same ROI for your 10 stacks over time you can then obtain measurements you can work with and plot.

Hope it helps.


Nguyen Phan
Biology Department
University of North Carolina at Chapel Hill
Chapel Hill, NC

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[Pablo Munoz-2]

thanks Nguyen , I will try to do it
El 09-07-2013, a las 0:24, Nguyen Phan <[hidden email]> escribió:

> Pablo:
>
> Since you are using a known cAMP probe, you can directly compare and plot the FRET intensity values at the neurite tip for each of the images. Alternately you can either look at the fluctuations of YFP or the YFP:CFP ratios for the ROI of interest. For more complicated measurements you can use normalized FRET which you dont necessarily have to use.
>
> I would use imageJ define the roi and obtain fl intensity measurements for them. Usng the same ROI for your 10 stacks over time you can then obtain measurements you can work with and plot.
>
> Hope it helps.
>
>
> Nguyen Phan
> Biology Department
> University of North Carolina at Chapel Hill
> Chapel Hill, NC
>
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> ImageJ mailing list: http://imagej.nih.gov/ij/list.html

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