Re: Assessing phototoxicity in live fluorescence imaging

Posted by Cole, Richard W (HEALTH) on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Re-Assessing-phototoxicity-in-live-fluorescence-imaging-tp7587005p7587013.html

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What I do when I REALLY need to measure/know power (not for the faint of heart).  Works on all imaging modalities, albeit w/some minor tweaks.  I detailed this in a paper which I can dig up an anyone is interested.

Basics: use match pair of immersion objectives configured such that the emitted light from one objective is collected by the 2nd and a power meter measures the light emitted from the 2nd objective.  I typical use a double coverslip dye sandwich for alignment and then remove for the final measurement.  
CLSM tweaks: bidirectional collection or park, zoom >2.5, longest dwell time/slowest scan speed
        Cautions: check all filter cubes/ AOTF settings
                   check illumination stability before starting


Richard Cole
Research Scientist V
Director: Advanced Light Microscopy & Image Analysis Core
Wadsworth Center
 
Research Assistant Professor
Dept. of Biomedical Sciences
School of Public Health State University of New York

120 New Scotland Avenue, Albany N.Y. 12208
518-474-7048 Phone
518-408-1730 Fax

Website http://www.wadsworth.org/research/cores/alm
 twitter.com/microscopejock