Large scale, single-cell FRET-based glucose uptake measurements within heterogeneous populations

Adam J M Wollman; Dimitrios Kioumourtzoglou; Rebecca Ward; Gwyn W Gould; Nia J Bryant

doi:10.1016/j.isci.2022.104023

Large scale, single-cell FRET-based glucose uptake measurements within heterogeneous populations

iScience. 2022 Mar 3;25(4):104023. doi: 10.1016/j.isci.2022.104023. eCollection 2022 Apr 15.

Authors

Adam J M Wollman^{1

2}, Dimitrios Kioumourtzoglou¹, Rebecca Ward¹, Gwyn W Gould³, Nia J Bryant¹

Affiliations

¹ Department of Biology and York Institute of Biomedical Research, University of York, York YO10 5DD, UK.
² Biosciences Institute, Newcastle University, Newcastle upon Tyne NE2 4HH, UK.
³ Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow G4 0RE, UK.

Abstract

Fluorescent biosensors are powerful tools allowing the concentration of metabolites and small molecules, and other properties such as pH and molecular crowding to be measured inside live single cells. The technology has been hampered by lack of simple software to identify cells and quantify biosensor signals in single cells. We have developed a new software package, FRETzel, to address this gap and demonstrate its use by measuring insulin-stimulated glucose uptake in individual fat cells of varying sizes for the first time. Our results support the long-standing hypothesis that larger fat cells are less sensitive to insulin than smaller ones, a finding that has important implications for the battle against type 2 diabetes. FRETzel has been optimized using the messy and crowded environment of cultured adipocytes, demonstrating its utility for quantification of FRET biosensors in a wide range of other cell types, including fibroblasts and yeast via a simple user-friendly quantitative interface.

Keywords: Biocomputational method; Bioinformatics; Biological sciences; Cell biology; Optical imaging.

Grants and funding

204829/WT_/Wellcome Trust/United Kingdom