Autofluorescence intensity imaging for studying mitochondrial dynamics

Blanche ter Hofstede; Amanda Galloway; Alex J. Walsh

doi:10.1117/12.3003064

12 March 2024 Autofluorescence intensity imaging for studying mitochondrial dynamics

Blanche ter Hofstede, Amanda Galloway, Alex J. Walsh

Author Affiliations +

Proceedings Volume 12854, Label-free Biomedical Imaging and Sensing (LBIS) 2024; 128540V (2024) https://doi.org/10.1117/12.3003064
Event: SPIE BiOS, 2024, San Francisco, California, United States

Conference Poster

Abstract

Mitochondria are highly dynamic organelles that continuously go through fission and fusion, a process that characterizes mitochondrial dynamics. These dynamics are important for the maintenance of the cell and are factors in aging, metabolic-related diseases, and cancer. Autofluorescence imaging (AFI) is used to study metabolic changes in the cell using the endogenous fluorophores reduced nicotinamide adenine dinucleotide (phosphate) NAD(P)H and flavin adenine dinucleotive (FAD). AFI can be used to study the dynamics of mitochondria, but requires high imaging speeds to capture the mitochondria movement. Here, we describe a multiphoton imaging technique that simultaneously captures NAD(P)H and FAD with a single excitation wavelength of 790 nm, reducing imaging times. A cyanide experiment was performed to verify that AFI at this optimal wavelength captures metabolic changes in cells. The optical redox ratios were computed from NAD(P)H and FAD images obtained both simultaneously using a single excitation wavelength and sequentially using the absorption-matched excitation wavelengths. The AFI results at 790 nm support the simultaneous acquisition of NAD(P)H and FAD for further research of mitochondrial dynamics and the metabolic state of the cell.

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

Citation Download Citation

Blanche ter Hofstede, Amanda Galloway, and Alex J. Walsh "Autofluorescence intensity imaging for studying mitochondrial dynamics", Proc. SPIE 12854, Label-free Biomedical Imaging and Sensing (LBIS) 2024, 128540V (12 March 2024); https://doi.org/10.1117/12.3003064

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