Differential levels of metabolic activity in isolated versus confluent/partially confluent HeLa cells are analyzed by autofluorescent NAD(P)H using multi-photon FLIM microscopy

Andrea Chandler; Aaron Chandler; Horst Wallrabe; Ammasi Periasamy

doi:10.1117/12.2267657

2 March 2017 Differential levels of metabolic activity in isolated versus confluent/partially confluent HeLa cells are analyzed by autofluorescent NAD(P)H using multi-photon FLIM microscopy

Andrea Chandler, Aaron Chandler, Horst Wallrabe, Ammasi Periasamy

Author Affiliations +

Proceedings Volume 10069, Multiphoton Microscopy in the Biomedical Sciences XVII; 100692R (2017) https://doi.org/10.1117/12.2267657
Event: SPIE BiOS, 2017, San Francisco, California, United States

Abstract

NAD(P)H is a known biomarker for cellular metabolism; a higher ratio of enzyme-bound NAD(P)H to free/unbound NAD(P)H indicates an increase in metabolic activity. Free NADH has a shorter fluorescence lifetime (τ₁), the bound version (τ₂) a longer lifetime. FLIM’s unique capability to establish inter alia the relative fractions of τ₁ (a₁%) and τ₂ (a₂%) in each pixel, determines the level of metabolic activity. The relative abundances of bound NAD(P)H were analyzed for single cells, confluent and partially confluent cells within 3 Fields-of-View (FoVs). A gradient of increasing a 2% levels of bound NAD(P)H from single, partially confluent to confluent cells was observed.

Citation Download Citation

Andrea Chandler, Aaron Chandler, Horst Wallrabe, and Ammasi Periasamy "Differential levels of metabolic activity in isolated versus confluent/partially confluent HeLa cells are analyzed by autofluorescent NAD(P)H using multi-photon FLIM microscopy", Proc. SPIE 10069, Multiphoton Microscopy in the Biomedical Sciences XVII, 100692R (2 March 2017); https://doi.org/10.1117/12.2267657

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