Multiphoton fluorescence lifetime imaging microscopy (FLIM) using a streak camera

Ramanujan Venkata Krishnan; Haruhisa Saitoh; Hirotoshi Terada; Victoria Frohlich Centonze; Brian Herman

doi:10.1117/12.478001

10 July 2003 Multiphoton fluorescence lifetime imaging microscopy (FLIM) using a streak camera

Ramanujan Venkata Krishnan, Haruhisa Saitoh, Hirotoshi Terada, Victoria Frohlich Centonze, Brian Herman

Author Affiliations +

Proceedings Volume 4963, Multiphoton Microscopy in the Biomedical Sciences III; (2003) https://doi.org/10.1117/12.478001
Event: Biomedical Optics, 2003, San Jose, CA, United States

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).

Citation Download Citation

Ramanujan Venkata Krishnan, Haruhisa Saitoh, Hirotoshi Terada, Victoria Frohlich Centonze, and Brian Herman "Multiphoton fluorescence lifetime imaging microscopy (FLIM) using a streak camera", Proc. SPIE 4963, Multiphoton Microscopy in the Biomedical Sciences III, (10 July 2003); https://doi.org/10.1117/12.478001

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