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Metabolic imaging by simultaneous FLIM of NAD(P)H and FAD
Author(s): Wolfgang Becker; Axel Bergmann; Rodrigo Suarez Ibarrola; Philippe-Fabian Müller; Lukas Braun
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Paper Abstract

We describe a metabolic-imaging system based on simultaneous recording of lifetime images of NAD(P)H and FAD. The system uses one-photon excitation by ps diode lasers, scanning by galvanometer mirrors, confocal detection, and two parallel TCSPC FLIM recording channels. The two lasers, with wavelengths of 375nm and 405 nm, are multiplexed to alternatingly excite NAD(P)H and FAD. One FLIM channel detects in the emission band of NAD(P)H, the other in the emission band of FAD. The FLIM data are processed by SPCImage data analysis software. For both channels, the data analysis delivers images of the amplitude-weighted lifetime, tm, the component lifetimes, t1 and t2, the amplitudes of the components, a1 and a2, and the amplitude ratio, a1/a2. Moreover, it delivers the fluorescence-lifetime redox ratio (FLIRR), a2nadh/a1fad. We demonstrate the performance of the system at the example of human bladder cells. Normal cells and tumor cells were discriminated by the tm images, the a1 images, and the FLIRR images.

Paper Details

Date Published: 22 February 2019
PDF: 8 pages
Proc. SPIE 10882, Multiphoton Microscopy in the Biomedical Sciences XIX, 108820B (22 February 2019); doi: 10.1117/12.2510132
Show Author Affiliations
Wolfgang Becker, Becker & Hickl GmbH (Germany)
Axel Bergmann, Becker & Hickl GmbH (Germany)
Rodrigo Suarez Ibarrola, Univ. of Freiburg – Medical Ctr. (Germany)
Philippe-Fabian Müller, Univ. of Freiburg – Medical Ctr. (Germany)
Lukas Braun, Becker & Hickl GmbH (Germany)


Published in SPIE Proceedings Vol. 10882:
Multiphoton Microscopy in the Biomedical Sciences XIX
Ammasi Periasamy; Peter T. C. So; Karsten König, Editor(s)

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