I just measured Fopt for my ‘scope

March 28, 2006 at 5:14 pm | | everyday science, hardware, tutorial

So here’s the deal: the detection efficiency for the stream of photons coming from one single fluorophore is kinda crappy (~10%). But that’s the state of the art; it’s part of the reason single-molecule measurements are so tough … and fun!

The detection efficiency D is:

D = nQFcollFoptFfilter

where nQ is the camera quantum efficiency, Fcoll is the percetage of light collected from the objective, Fopt is the efficiency through all the optics in the microscope, and Ffilter is the transmission through the various filters. Today, I measured Fopt to be 50% at 488 nm. Here’s a page from my notebook that shows my setup:

p73pic

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