Differences
This shows you the differences between two versions of the page.
Both sides previous revisionPrevious revisionNext revision | Previous revisionNext revisionBoth sides next revision |
howto:flim-fret_calculation_for_single_exponential_donors [2014/05/23 07:43] – admin | howto:flim-fret_calculation_for_single_exponential_donors [2014/06/18 09:16] – admin |
---|
| |
This tutorial shows step-by-step, how the "Lifetime FRET Image" analysis of [[products:SymPhoTime64|SymPhoTime 64]] can be used to calculate pixel-by-pixel the FRET efficiency in an image containing a cell transfected with a [[wp>Green_fluorescent_protein|GFP]] and a RFP construct, that bind in certain regions of the cytoplasm. A bi-exponential fit is performed in a larger region of interest to determine the donor only lifetime and the lifetime of the donor in presence of [[glossary:fret|FRET]]. A pixel-by-pixel fit is performed in the image and in every pixel the FRET efficiency as well as the fraction of FRET on the total decay ("Binding") is determined. | This tutorial shows step-by-step, how the "Lifetime FRET Image" analysis of [[products:SymPhoTime64|SymPhoTime 64]] can be used to calculate pixel-by-pixel the FRET efficiency in an image containing a cell transfected with a [[wp>Green_fluorescent_protein|GFP]] and a RFP construct, that bind in certain regions of the cytoplasm. A bi-exponential fit is performed in a larger region of interest to determine the donor only lifetime and the lifetime of the donor in presence of [[glossary:fret|FRET]]. A pixel-by-pixel fit is performed in the image and in every pixel the FRET efficiency as well as the fraction of FRET on the total decay ("Binding") is determined. |
| |
| {{ youtube>Ka51hZNw3u8?large }} |
| |
===== Background information ===== | ===== Background information ===== |