Fluorescence is emitted from a single fluorophore with a lifetime of τ. The idealized exponential decay is indicated by the dottedline in Figure
1. However, you have a detector with a signifcant exposure time ?t, such that the fluorescence level changes during the exposure. Your setup allows you to trigger two such exposures D1 and D2, indicated in the figure. As an experimenter, you know the two measurements began (were triggered) at 0.8 and 2.5 nsec, but you do not know ?t, the exposure time (the areas in the figure are just for illustration).
You have a linear array of 51 such detectors. On the website, you can download a file called FLIMdata.dat, a 2-by-51 matrix that represents D1 and D2 measurements at each detector. Plots of the D1 and D2 intensities versus detector position (1 through 51) are shown in Figure 2. Your job is to make a plot of τ , the fluorescence lifetime, versus detector position (which would correspond to, for example, a linear cut across a region of cells).
This will require some open-ended thinking about how to extract τ from the measured data. You should find that the calculated lifetime is much more spatially uniform than the noisy intensity data, with a single well-defined region of spatial contrast. What is the uniform background lifetime, in nanoseconds, and what is the lifetime in the contrasting region?
Attachment:- Flim.m.txt