Time-gated luminescence microscopy.
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 micros 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.
Connally
Russell E
RE
Centre for Laser Applications, Department of Physics, Division of Information and Communication Sciences, Macquarie University, Sydney, NSW 2109, Australia. rconnall@ics.mq.edu.au
Piper
James A
JA
D000818:Animals; D004867:Equipment Design; D056890:Eukaryota / Q000378:metabolism*; D005872:Giardia; D007091:Image Processing, Computer-Assisted; D028581:Lanthanoid Series Elements / Q000737:chemistry; D007834:Lasers; D008027:Light; D049449:Luminescence; D008853:Microscopy / Q000295:instrumentation / Q000379:methods*; D008860:Microscopy, Ultraviolet / Q000379:methods; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013997:Time Factors; D014466:Ultraviolet Rays
D028581:Lanthanoid Series Elements