Protocol for pH correction of FRET-based fluorescent biosensor imaging in dissociated Drosophila neurons.

Authors

Matthew S Price, Elham Rastegari, Kartik Venkatachalam

Published in

STAR protocols. Volume 7. Issue 3. Pages 104802. Aug 27, 2026. Epub Aug 27, 2026.

Abstract

Fluorescent biosensors are sensitive to intracellular pH fluctuations, which can shift fluorophore properties and compromise accurate metabolite quantification. We present a protocol for pH correction in FRET-based metabolic biosensor imaging using dissociated Drosophila larval neurons. We describe steps for integrating cytosolic pH measurements with R-based post hoc correction of FRET ratios. We also detail procedures for brain dissection, tissue dissociation, primary neuron culture, and live-cell imaging. This protocol enables accurate metabolic biosensor readouts at single-cell resolution. For complete details on the use and execution of this protocol, please refer to Price et al.¹.

PMID:
42664056
Bibliographic data and abstract were imported from PubMed on 29 Aug 2026.

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