Fluorescence Lifetime Imaging Microscope Consisting of a Compact Picosecond Dye Laser and a Gated Charge-Coupled Device Camera for Applications to Living Cells

Abstract

An inverted microscope was combined with a compact dye laser with a pulse width of <190 ps and an intensified charge-coupled device (ICCD) camera with a minimum gate width of 200 ps. The resulting fluorescence lifetime imaging microscope, which has a temporal resolution of 340 ps, was used to measure the fluorescence lifetime of polymer microspherers. The results indicated a fluorescence lifetime of 0.9 ns. The present analytical instrument was also employed in an evaluation of biological cells after labeling them with SYTO 13, a fluorescent dye.