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Proceedings Paper

Multiphoton fluorescence lifetime imaging microscopy (FLIM) using a streak camera
Author(s): Ramanujan Venkata Krishnan; Haruhisa Saitoh; Hirotoshi Terada; Victoria Frohlich Centonze; Brian Herman
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Paper Abstract

We report the development, calibration and biomedical application of a multiphoton fluorescence lifetime imaging system (FLIM) using a streak camera. The present system is versatile with high spatial (approximately 0.2 μm) and temporal (approximately 50 psec) resolution and allows rapid data acquisition and reliable and reproducible lifetime determinations. The system was calibrated with standard fluorescent dyes and the lifetime values obtained were in very good agreement with values reported in literature for these dyes. We also demonstrate the applicability of the system to FLIM studies in cellular specimens in the context of quantitative measurement of fluorescence resonance energy transfer (FRET).

Paper Details

Date Published: 10 July 2003
PDF: 10 pages
Proc. SPIE 4963, Multiphoton Microscopy in the Biomedical Sciences III, (10 July 2003); doi: 10.1117/12.478001
Show Author Affiliations
Ramanujan Venkata Krishnan, Univ. of Texas Health Science Ctr. (United States)
Haruhisa Saitoh, Hamamatsu Photonics KK (Japan)
Hirotoshi Terada, Hamamatsu Photonics KK (Japan)
Victoria Frohlich Centonze, Univ. of Texas Health Science Ctr. (United States)
Brian Herman, Univ. of Texas Health Science Ctr. (United States)

Published in SPIE Proceedings Vol. 4963:
Multiphoton Microscopy in the Biomedical Sciences III
Ammasi Periasamy; Peter T. C. So, Editor(s)

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