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Methods Enzymol. 2006;414:633-42.

Fluorescence lifetime imaging microscopy: two-dimensional distribution measurement of fluorescence lifetime.

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1
Hamamatsu Photonic Systems, Bridgewater, NJ, USA.

Abstract

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.

PMID:
17110215
DOI:
10.1016/S0076-6879(06)14033-1
[Indexed for MEDLINE]
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