4.7 Article Proceedings Paper

Time-gated luminescence microscopy

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BLACKWELL PUBLISHING
DOI: 10.1196/annals.1430.032

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time-gated luminescence; autofluorescence; lanthanide; phosphorescence; Giardia; EMCCD; UV LED

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Autofluorescent algal samples were spiked with europium. beads for analysis on a novel all-solid-state, time-gated luminescence (TGL) microscope. Pulsed UV excitation (365 nm) was provided by a high-power UV-LED source fitted to an Olympus BX51 microscope. An Impactron electron multiplying charge-coupled-device (CCD) camera acquired images in delayed luminescence mode. Second, we evaluated sensitivity of the instrument by acquiring images of immunofluorescently labeled Giardia cysts with a single-exposure period of 3 ms. The camera was triggered 3 mu s after the LED had extinguished to yield a 14-fold increase in signal-to-noise ratio within a single 33 ms capture cycle. This novel instrument could be switched instantly from prompt epifluorescence mode to TGL mode for suppression of short-lived fluorescence.

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