Re: TIRF

Posted by Mancini, Michael A on
URL: http://confocal-microscopy-list.275.s1.nabble.com/reducing-size-of-illumination-in-epi-fluorescence-tp593586p593596.html

Speaking of TIRF issues, perhaps list members would like to comment on some differing opinions I've been hearing about the usefulness of a TIRF lens for standard fluorescence. I've heard two extremes on the appropriateness/usefulness of a 1.49 NA TIRF lens for high mag/high res/high sensitivity imaging away from the substratum (for example, intranuclear structures). Although I'm a bit physics-challenged, it seems to make sense to me that the NA would not be realized in a glycerol mounting medium due to RI mismatch, and could be counter productive, in fact. Seems the vendors that sell TIRF lens-systems say they are great for every use, pumping up the high NA as a cure for everything, and vendors that don't sell them say they're only good for a true TIRF application.

Comments are welcome both on/off the list.

Cheers,

Mike
_______________________
Michael A. Mancini, Ph.D
Department of Molecular and Cellular Biology
Baylor College of Medicine
Houston, TX 77030
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713.408.0179 cell