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Proceedings Paper

Picosecond fluorescence lifetime imaging microscope for imaging of living glioma cells
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Paper Abstract

In this communication, we report the imaging of living glioma cells using fluorescence lifetime imaging (FLIM) technique. The growing interests in developing novel techniques for diagnosis and minimally invasive therapy of brain tumor have led to microscopic studies of subcellular structures and intracellular processes in glioma cells. Fluorescence microscopy has been used with a number of exogenous molecular probes specific for certain intracellular structures such as mitochondria, peripheral benzodiazepine receptor (PBR), and calcium concentration. When probes with overlapping emission spectra being used, separate samples are required to image each probe individually under conventional fluorescence microscopy. We have developed a wide-field FLIM microscope that uses fluorescence lifetime as an additional contrast for resolving multiple markers in the same essay. The FLIM microscope consists of a violet diode laser and a nitrogen-pumped dye laser to provide tunable sub-nanosecond excitation from UV to NIR. The detection system is based on a time-gated ICCD camera with minimum 80 ps gate width. The performance of the system was evaluated using fluorescence dyes with reported lifetime values. Living rat glioma C6 cells were stained with JC-1 and Rhodamine 123. FLIM images were acquired and their lifetimes in living cells were found in good agreements with values measured in solutions by a time-domain fluorescence spectrometer. These results indicate that imaging of glioma cells using FLIM can resolve multiple spectrally-overlapping probes and provide quantitative functional information about the intracellular environment.

Paper Details

Date Published: 29 March 2005
PDF: 7 pages
Proc. SPIE 5699, Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III, (29 March 2005); doi: 10.1117/12.592474
Show Author Affiliations
Qiyin Fang, Cedars-Sinai Medical Ctr. (United States)
Jingjing Wang, Univ. of Southern California (United States)
Yinghua Sun, Univ. of Southern California (United States)
Thomas Vernier, Univ. of Southern California (United States)
Thanassis Papaioannou, Cedars-Sinai Medical Ctr. (United States)
Javier Jo, Cedars-Sinai Medical Ctr. (United States)
Mya Mya Thu, Univ. of Southern California (United States)
Martin A. Gundersen, Univ. of Southern California (United States)
Laura Marcu, Cedars-Sinai Medical Ctr. (United States)
Univ. of Southern California (United States)


Published in SPIE Proceedings Vol. 5699:
Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III
Dan V. Nicolau; Ramesh Raghavachari; Dan V. Nicolau; Jörg Enderlein; Robert C. Leif; Daniel L. Farkas, Editor(s)

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