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Proceedings Paper

Metabolic imaging by simultaneous 2-photon FLIM of NAD(P)H and FAD
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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 two-photon excitation by a dual-wavelength femtosecond fibre laser. The two wavelengths of the laser, 780 nm and 880 nm, are multiplexed synchronously with the frames or the lines of the scan. The recording system uses two parallel TCSPC FLIM channels, detecting from 420 to 475 nm and 480 to 600 nm. By using the multiplexing functions of the TCSPC modules, separate images for NAD(P)H and FAD are recorded. A third image is obtained for the SHG of the 880 nm laser wavelength. Data analysis delivers images of the amplitude-weighted lifetime, tm, the component lifetimes, t1 and t2, the amplitudes of the components, a1 and a2, the amplitude ratio, a1/a2, and the fluorescence-lifetime redox ratio (FLIRR), a2nadh/a1fad. We demonstrate the performance of the system for metabolic imaging of mammalian skin.

Paper Details

Date Published: 14 February 2020
PDF: 5 pages
Proc. SPIE 11244, Multiphoton Microscopy in the Biomedical Sciences XX, 112440L (14 February 2020);
Show Author Affiliations
Wolfgang Becker, Becker & Hickl GmbH (Germany)
Axel Bergmann, Becker & Hickl GmbH (Germany)
Alexander Jelzow, Becker & Hickl GmbH (Germany)
Antje Neubauer, Becker & Hickl GmbH (Germany)
Angelika Rück, Univ. Ulm (Germany)
Konrad Birkmeier, TOPTICA Photonics AG (Germany)
Patrick Leisching, TOPTICA Photonics AG (Germany)

Published in SPIE Proceedings Vol. 11244:
Multiphoton Microscopy in the Biomedical Sciences XX
Ammasi Periasamy; Peter T. C. So; Karsten König, Editor(s)

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