Journal
OPTICS LETTERS
Volume 39, Issue 19, Pages 5602-5605Publisher
OPTICAL SOC AMER
DOI: 10.1364/OL.39.005602
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Funding
- MRC [G0802216] Funding Source: UKRI
- Engineering and Physical Sciences Research Council [EP/C510135/1] Funding Source: researchfish
- Medical Research Council [G0802216] Funding Source: researchfish
- Medical Research Council [G0802216] Funding Source: Medline
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A 1 MHz frame rate complementary metal-oxide semiconductor (CMOS) camera was used in combination with an image intensifier for wide-field time-correlated single-photon counting (TCSPC) imaging. The system combines an ultrafast frame rate with single-photon sensitivity and was employed on a fluorescence microscope to image decays of ruthenium compound Ru(dpp) with lifetimes from around 1 to 5 mu s. A submicrowatt excitation power over the whole field of view is sufficient for this approach, and compatibility with live-cell imaging was demonstrated by imaging europium-containing beads with a lifetime of 570 mu s in living HeLa cells. A standard two-photon excitation scanning fluorescence lifetime imaging (FLIM) system was used to independently verify the lifetime for the europium beads. This approach brings together advantageous features for time-resolved live-cell imaging such as low excitation intensity, single-photon sensitivity, ultrafast camera frame rates, and short acquisition times. (C) 2014 Optical Society of America
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