Přejít k obsahu
Merck

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

Molecular metabolism (2024-07-07)
Ariful Haque Abir, Leonie Weckwerth, Artur Wilhelm, Jana Thomas, Clara M Reichardt, Luis Munoz, Simon Völkl, Uwe Appelt, Markus Mroz, Raluca Niesner, Anja Hauser, Rebecca Sophie Fischer, Katharina Pracht, Hans-Martin Jäck, Georg Schett, Gerhard Krönke, Dirk Mielenz
ANOTACE

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. 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. 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 naïve 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 naïve and memory B-lymphocytes. Taken together, these data show that the optical redox ratio can depict metabolic differences in distinct cell populations by flow cytometry.

MATERIÁLY
Číslo produktu
Značka
Popis produktu

Sigma-Aldrich
Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone, ≥98% (TLC), powder
Sigma-Aldrich
2-Deoxy-D-glucose, ≥98% (GC), crystalline
Sigma-Aldrich
Rotenone, ≥95%
Sigma-Aldrich
Nicotinamide Phosphoribosyltransferase Inhibitor, FK866, The Nicotinamide Phosphoribosyltransferase Inhibitor, FK866, also referenced under CAS 658084-64-1, controls the biological activity of Nicotinamide Phosphoribosyltransferase. This small molecule/inhibitor is primarily used for Neuroscience applications.