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Publication : Metabolic profiling of single cells by exploiting NADH and FAD fluorescence via flow cytometry.

First Author  Abir AH Year  2024
Journal  Mol Metab Volume  87
Pages  101981 PubMed ID  38971403
Mgi Jnum  J:372515 Mgi Id  MGI:7708427
Doi  10.1016/j.molmet.2024.101981 Citation  Abir AH, et al. (2024) Metabolic profiling of single cells by exploiting NADH and FAD fluorescence via flow cytometry. Mol Metab 87:101981
abstractText  OBJECTIVE: The metabolism of different cells within the same microenvironment can differ and dictate physiological or pathological adaptions. Current single-cell analysis methods of metabolism are not label-free. METHODS: The study introduces a label-free, live-cell analysis method assessing endogenous fluorescence of NAD(P)H and FAD in surface-stained cells by flow cytometry. RESULTS: OxPhos inhibition, mitochondrial uncoupling, glucose exposure, genetic inactivation of glucose uptake and mitochondrial respiration alter the optical redox ratios of FAD and NAD(P)H as measured by flow cytometry. Those alterations correlate strongly with measurements obtained by extracellular flux analysis. Consequently, metabolically distinct live B-cell populations can be resolved, showing that human memory B-cells from peripheral blood exhibit a higher glycolytic flexibility than naive B cells. Moreover, the comparison of blood-derived B- and T-lymphocytes from healthy donors and rheumatoid arthritis patients unleashes rheumatoid arthritis-associated metabolic traits in human naive and memory B-lymphocytes. CONCLUSIONS: Taken together, these data show that the optical redox ratio can depict metabolic differences in distinct cell populations by flow cytometry.
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