Metabolic imaging by simultaneous 2-photon FLIM of NAD(P)H and FAD

Wolfgang Becker; Axel Bergmann; Alexander Jelzow; Antje Neubauer; Angelika Rück; Konrad Birkmeier; Patrick Leisching

doi:10.1117/12.2550962

14 February 2020 Metabolic imaging by simultaneous 2-photon FLIM of NAD(P)H and FAD

Wolfgang Becker, Axel Bergmann, Alexander Jelzow, Antje Neubauer, Angelika Rück, Konrad Birkmeier, Patrick Leisching

Proceedings Volume 11244, Multiphoton Microscopy in the Biomedical Sciences XX; 112440L (2020) https://doi.org/10.1117/12.2550962
Event: SPIE BiOS, 2020, San Francisco, California, United States

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), a_2nadh/a_1fad. We demonstrate the performance of the system for metabolic imaging of mammalian skin.

Conference Presentation

Citation Download Citation

Wolfgang Becker, Axel Bergmann, Alexander Jelzow, Antje Neubauer, Angelika Rück, Konrad Birkmeier, and Patrick Leisching "Metabolic imaging by simultaneous 2-photon FLIM of NAD(P)H and FAD", Proc. SPIE 11244, Multiphoton Microscopy in the Biomedical Sciences XX, 112440L (14 February 2020); https://doi.org/10.1117/12.2550962

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available