I have been using AccPbFRET to analyze my FRET data. I used to use ivision, which is too expensive. This software is free, and I have tested it with my old samples, and it proved to be OK, generating some consistent data. However, I am confused about the way that it took to get the mean FRET value from the FRET map. What is the "not NaN P.?" Does it count every pixel that I select or only the "not NaN P."? The FRET value also seems different then when I use high threshold to only highlight the cell membrane part and get FRET v.s. when I use low threshold >20 not to highlight the membrane, but used a ROI to select membrane part, and get FRET. Would these two ways of analysis give me different results?
Thank you so much!
In attached paper is example of FRET between fluorescent protein and conducting polymer.
Article Fluorescence study of glucose oxidase self-encapsulated with...
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