Posted by
Alessandro Esposito on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Several-Alexa-Fluor-dyes-extinction-coefficients-and-quantum-yields-tabulated-new-deconvolution-papes-tp7581205p7581235.html
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Dear Gabor,
you may find some information in Qiaole Zhang et al published in J Biomed
Opt "Photon budget analysis for fluorescence lifetime imaging microscopy"
As you said, a major impact is the NA of the objective. For single molecule
detection using top end EM-CCD, you will not lose much at the camera.
However, you will lose quite a bit into the filters and relay optics. I've written
a short description about this in Suppoprting Text S1 here:
<a href="http://www.plosone.org/article/info%3Adoi%2F10.1371%">http://www.plosone.org/article/info%3Adoi%2F10.1371%
2Fjournal.pone.0077392
Cheers,
Alessandro
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>Dear All,
>
>I made a quick search but found nothing really useful. My question is the
>following: does anyone have some data/literature how efficient microscopes
>are in detecting the photons of a fluorescent sample? Obviously the NA and
>the QE of the camera (plus the performance filters/dichroics) are
>important factors but taken e.g. A single molecule experiment with a 1.49
>NA objective and a back-illuminated EM-CCD what ration of the emitted
>photons really reach the detector? Of course the transmission of the
>objectives and other lenses in the microscopes can vary but is there still
>an estimate?
>
>Thanks Gabor