Light Source Illumination at the Microscope Stage

Autofluorescent slides were used to quantify the brightness at the stage of a Zeiss Axioskop using a 20 X plan-ACHROMAT objective, a Quantix camera, and the grey level function of MetaMorph. In each case, five separate measurements were taken from different parts of the slide. Data represent mean +/- standard deviation. For DAPI/UV measurements a 49000 filter set was used (excitation D350/50x) with a blue autofluorescent slide, and the exposure time was 25 ms. For FITC/GFP measurements, a 49002 set was used (excitation ET470/40x) with a 1.3 ND neutral density filter, a red autofluorescent slide and an exposure time of 400 ms. For TRITC/Cy3 measurements, a 49004 filter set was used (excitation ET545/25x) with a 1.3 ND neutral density filter, a yellow autofluorescent slide, and an exposure time of 80 ms. For Cy5 measurements, a 49006 filter set was used (excitation ET620/60x) with a 1.3 ND neutral density filter, a red autofluorescent slide, and the exposure time was 240 ms. Lifetime lamp use time was 42 h, 8 h, and 89 h for the PhotoFluor® II, mercury lamp, and 120 W metal halide, respectively.

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Output power in the ultraviolet through visible spectrum

Comparison of the PhotoFluor II lamp (green trace) and a 120 W metal halide lamp (orange trace). Output from the end of the liquid light guide was measured using a spectrometer.