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.
cenarios are illustrated. In <imgref figure1>A on average only 1 fluorescent molecule resides in the detect... y when the molecule is in) in comparison with the average value. In <imgref figure1>B a few molecules reside in the detection volume on average, and thus the relative fluctuations will be small... re a few molecules in focus) in comparison to the average. This difference in the relative fluctuations is
erential count rate//. That means NOT at the same average count rate. //Differential count rate// is the re... ). However, for an IRF measurement the same "low" average count rate can be far too high.
Consider a 150 ps FWHM IRF measured at an average count rate of 100 kcps (kilocounts per second) at... till just an optimistic approximation based on an average value over the whole pulse duration. At the momen
aking room for fresh incoming events. Even if the average read rate of the host PC is limited, bursts with ... r rate can be recorded for some time. Only if the average count rate over a long period of time exceeds the... r, on a modern and well configured PC a sustained average count rate over 4 Mcps is possible. This total tr... efficient processing and possible restrictions in average count rate. The PicoHarp software installation CD
aking room for fresh incoming events. Even if the average read rate of the host PC is limited, bursts with ... r rate can be recorded for some time. Only if the average count rate over a long period of time exceeds the... configured PC with Windows 2000 or XP a sustained average count rate over 4 Mcps is possible. This total tr... efficient processing and possible restrictions in average count rate. The *Harp software installation CD co
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
Fast lifetime ======
A method for estimating the average lifetime, calculated non-distinctive on different... the IRF to the barycentre of the decay equals the average lifetime. This estimate is very fast and does not... he [[FWHM]] interval. However, this may introduce a systematic shift in the estimated average lifetimes.
he optical system is randomly changing around the average number. The analysis gives the average number of fluorescent particles and average diffusion time, when the particle is passing through the space
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
TCSPC devices. There are two effects:
* The average measured lifetime becomes shorter
* A mono-expo... gram the following results are obtained:
* the average lifetime decrease is 3%
* the χ2 value increase
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