In vivo two-photon FLIM imaging to investigate metabolism in live animals

Vijay Kumar Sagar; Andrés Norambuena; Horst Wallrabe; Shagufta R. Alam; Evelyn Pardo; Ammasi Periasamy

doi:10.1117/12.3003082

12 March 2024 In vivo two-photon FLIM imaging to investigate metabolism in live animals

Vijay Kumar Sagar, Andrés Norambuena, Horst Wallrabe, Shagufta R. Alam, Evelyn Pardo, Ammasi Periasamy

Proceedings Volume 12847, Multiphoton Microscopy in the Biomedical Sciences XXIV; 128470Q (2024) https://doi.org/10.1117/12.3003082
Event: SPIE BiOS, 2024, San Francisco, California, United States

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Abstract

The advances in the field of optics and laser technologies are helpful to visualize and investigate the metabolism in live animals. Fluorescence lifetime imaging (FLIM) is a non-invasive optical technique to measure the fluorescence lifetime of the fluorophore towards its applications where fluorescence intensity-based techniques do not provide sufficient and accurate information to discriminate auto-fluorescent species. In contrast to conventional single photon excitation, two-photon excitation improves the penetration depth with low scattering of the longer wavelength to examine thick biological samples. In this work, we explain our 2photon-FLIM methodology for imaging mouse brains via a cranial window to investigate the changes in the NAD(P)H bound coenzyme, in both wild type and Alzheimer disease (AD) animal models before and after stimulation with nutrients. Moreover, phasor plot analysis has been used for removing lower a₂(%) fraction from the metabolic trajectory.

(2024) Published by SPIE. Downloading of the abstract is permitted for personal use only.

Citation Download Citation

Vijay Kumar Sagar, Andrés Norambuena, Horst Wallrabe, Shagufta R. Alam, Evelyn Pardo, and Ammasi Periasamy "In vivo two-photon FLIM imaging to investigate metabolism in live animals", Proc. SPIE 12847, Multiphoton Microscopy in the Biomedical Sciences XXIV, 128470Q (12 March 2024); https://doi.org/10.1117/12.3003082

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