I have a fluorescence microscopy image of nanoparticles and I wish to measure the quantum yield of nanoparticles and the fluorescence decay of nanoparticles. However I have no idea how to proceed and which software to use.
If you have experience in interpreting fluorescence microscopy images and its analysis please provide some guidance on how to proceed.
if you have fluorescent NP then why donot you try spectroscopically instead of microscopically
Unfortunately,there is only one corner stone, calibrations : this kind of problem requires to plot a curve "calibrate the grey levels of the image pixels vs incident energy --for each pertinent energy band-- of a calibrated excitation source".
I would record the same image at different excitation energies. If you plot then the average pixel intensity versus excitation energy then you may derive the slope from it.
But you have to substract also the ground effect. On the same graph you have to plot the pixel intensity for a region where you don't have nanoparticles.
If you do like this for many nanoparticles (the same nanoparticles) then you may get an average quantum yield.
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