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A streamlined spectral cytometry method for FAD and NADH autofluorescence analysis in immunometabolic studies

Preprint Created on 09 Jun 2026 bioRxiv

We present a streamlined protocol that enables the characterization of the metabolic state of immune cell populations through their distinct NADH/FAD autofluorescence fingerprints using a FACSymphony A5 spectral cytometer. We demonstrate the utility of this approach by profiling the metabolic status of diverse splenic B-cell subsets and assessing metabolic changes associated with their activation state.

Stylianakis, E., Hoevelmeyer, N.

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