Multiphoton fluorescence lifetime imaging of NADH reveals spatio-temporal patterns in cell metabolism during collective migration

Jacopo Ferruzzi; Stephen J. DeCamp; Victor M. K. Tsuda; Muhammad H. Zaman; Jeffrey J. Fredberg; Darren M. Roblyer

doi:10.1117/12.2578743

5 March 2021 Multiphoton fluorescence lifetime imaging of NADH reveals spatio-temporal patterns in cell metabolism during collective migration

Jacopo Ferruzzi, Stephen J. DeCamp, Victor M. K. Tsuda, Muhammad H. Zaman, Jeffrey J. Fredberg, Darren M. Roblyer

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Proceedings Volume 11622, Multiscale Imaging and Spectroscopy II; 116220G (2021) https://doi.org/10.1117/12.2578743
Event: SPIE BiOS, 2021, Online Only

Abstract

Fluorescence lifetime imaging (FLIM) is a powerful tool to quantify local changes in cell metabolism without loss of spatial resolution. Here, we adopted FLIM to characterize spatio-temporal metabolic changes occurring during collective cell migration. Using an established in vitro system, we measured biomechanical and metabolic changes during migration of epithelial cell monolayers. We developed a custom image analysis pipeline that combines machine learning segmentation and curve fitting analysis to analyze FLIM data. Our findings – which were validated via separate measurements of cytoplasmic redox ratio, glucose uptake, and mitochondrial membrane potential – are consistent with a glycolytic shift during collective cell migration.

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Jacopo Ferruzzi, Stephen J. DeCamp, Victor M. K. Tsuda, Muhammad H. Zaman, Jeffrey J. Fredberg, and Darren M. Roblyer "Multiphoton fluorescence lifetime imaging of NADH reveals spatio-temporal patterns in cell metabolism during collective migration", Proc. SPIE 11622, Multiscale Imaging and Spectroscopy II, 116220G (5 March 2021); https://doi.org/10.1117/12.2578743

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KEYWORDS

Mode conditioning cables

Multiphoton fluorescence microscopy

Image analysis

Image resolution

In vitro testing

In vivo imaging

Multiphoton microscopy

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