Advancing optical metabolic imaging through phasor-based S-FLIM-SHG microscopy

Belen Torrado; Alexander Vallmitjana; Michelle A. Digman; Alexander Dvornikov; Enrico Gratton

doi:10.1117/12.3009991

13 March 2024 Advancing optical metabolic imaging through phasor-based S-FLIM-SHG microscopy

Belen Torrado, Alexander Vallmitjana, Michelle A. Digman, Alexander Dvornikov, Enrico Gratton

Proceedings Volume PC12847, Multiphoton Microscopy in the Biomedical Sciences XXIV; PC128470R (2024) https://doi.org/10.1117/12.3009991
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

This study addresses challenges in Optical Metabolic Imaging due to dim signals, overlapping spectra, and similar lifetimes of NADH and FAD autofluorescent molecules. A Phasor-based S-FLIM-SHG microscope is introduced, enabling simultaneous acquisition of Hyperspectral Imaging (HSI), Fluorescence Lifetime Imaging Microscopy (FLIM), and Second Harmonic Generation imaging (SHG). The microscope's design efficiently detects scattered photons from complex samples and it is particularly competent at detection SHG signal. A novel 5D-snapshot (x, y, z, τ, λ). metabolic imaging method is proposed, significantly reducing acquisition times and enhancing measurement accuracy. The method's versatility is demonstrated across diverse sample types, with potential implications for advancing optical metabolic imaging capabilities.

Conference Presentation

Citation Download Citation

Belen Torrado, Alexander Vallmitjana, Michelle A. Digman, Alexander Dvornikov, and Enrico Gratton "Advancing optical metabolic imaging through phasor-based S-FLIM-SHG microscopy", Proc. SPIE PC12847, Multiphoton Microscopy in the Biomedical Sciences XXIV, PC128470R (13 March 2024); https://doi.org/10.1117/12.3009991

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