me 64 can be used to calculate pixel-by-pixel the average [[glossary:fret|FRET]] efficiency in an image con... ed with Cerulean/YFP-constructs. To calculate the average FRET-efficiency, first the donor only lifetime is... it with the FLIM-analysis. The amplitude weighted average donor lifetime is used to calculate the average FRET-efficiency using the "Lifetime FRET Image" analysis.
gram the following results are obtained:
* the average lifetime decrease is 3%
* the $\chi^2$ value in... of Points" should be set to 100. In this way, the average count rate is much more realistic especially for ... xels. This might not be suited to set the correct average count rate.))
* At a pixel dwell time of 10µs e... the time trace. At a pixel dwell time of 1 µs the average of 10 pixels are displayed as one point in the ti
2D histogram. The x-axis shows a histogram of the average lifetimes of the pixels in the image. The y-axis ... 00 }}
**Note:** In the example, two peaks in the average lifetime (x-axis) can already be seen.
* Zoom ... Fitting feature and calculating the values of the average FRET efficiency and binding (not shown).
==== Pe... l_fit.png}} feature decomposes the overall decay (average over all pixels) into the user-defined patterns.
ncodes the intensity, while the color encodes the average "Fast FLIM" lifetime, i.e. the mean arrival times... the dark background is usually very long (with an average up to 1/2 of the TCSPC window), which makes the c... change of the fit values and compare for instance average lifetime of your ROIs.
* Select the intensity ... lot_10framebinning_2.png |}}
**Response:**\\
The average lifetime, you can see that after adding the sugar
64]] can be used to calculate pixel-by-pixel the average [[glossary:fret|FRET]] efficiency in an image con... ed with Cerulean/YFP constructs. To calculate the average FRET efficiency, a ratiometric approach based on ... y FRET as used here can only be used to calculate average FRET efficiencies. In case of a mixed population,
encodes the intensity while the color encodes the average "Fast FLIM" lifetime, i.e. the mean arrival times... s sufficient for the model suggested, click on an average pixel of interest in the FLIM image.
**Response:... change, as now the calculated intensity weighted average lifetime "tau_Av_Int" is displayed, in contrast t
ncodes the intensity, while the color encodes the average "Fast FLIM" lifetime, i.e. the mean arrival times... the dark background is usually very long (with an average up to 1/2 of the TCSPC window), which makes the c... change, as now the calculated intensity weighted average lifetime "tau_Av_Int" is displayed, in contrast t
encodes the intensity while the color encodes the average "Fast FLIM" lifetime, i.e. the mean arrival time ... tor, the binding ratio has a direct effect on the average lifetime.
{{ :howto:flim-fret-1expd_calcfret_resp
ncodes the intensity, while the color encodes the average "Fast FLIM" lifetime, i.e. the mean arrival times... the dark background is usually very long (with an average up to 1/2 of the TCSPC window), which makes the c
encodes the intensity while the color encodes the average "Fast FLIM" lifetime, i.e. the mean arrival times... to display (e.g. T_av_Int, the intensity-weighted average lifetime, which - neglecting background photons –
* G(0)
* Num. Molecules (calculated from the average countrate and G(0))
* mol. Brightness (in kcps/... ideal system should equal the intensity weighted average lifetime [τ av. int.]. Usually, fitted lifetimes
ncodes the intensity, while the color encodes the average "Fast FLIM" lifetime, i.e. the mean arrival times... the dark background is usually very long (with an average up to 1/2 of the TCSPC window), which makes the c