Fluorescence lifetime imaging microscopy: two-dimensional distribution measurement of fluorescence lifetime.
A newly developed fluorescence lifetime imaging microscopy (FLIM) system has combined the high-temporal resolution of a streak camera with the high-spatial resolution of a microscope to obtain a two-dimensional distribution of fluorescence lifetimes within living cells. The temporal resolution is as short as 20 ps. The effective field of view is 48 x 45 mum with 0.2 mum resolution using a 60x water immersion objective. Image acquisition time is as short as 3 s per image. Measured and published values of lifetime for standard fluophores are shown with good agreement. Examples of FLIM and fluorescence resonance energy transfer images are presented.
Fujiwara
Masanobu
M
Hamamatsu Photonic Systems, Bridgewater, NJ, USA.
Cieslik
William
W
D000818:Animals; D002138:Calibration; D002460:Cell Line; D006224:Cricetinae; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes / Q000494:pharmacology; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D012107:Research Design; D013997:Time Factors
D005456:Fluorescent Dyes