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
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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.
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Andrea Chandler, Andrea Chandler, Aaron Chandler, Aaron Chandler, Horst Wallrabe, Horst Wallrabe, Ammasi Periasamy, 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); doi: 10.1117/12.2267657; https://doi.org/10.1117/12.2267657
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