Pulmonary/Critical Care Research

Single-cell lactate production rate as a measure of glycolysis in endothelial cells

Devin Harrison,1,2,3,4 David Wu,2,4,5 Jun Huang,1,3,* and Yun Fang1,2,6,**

1Graduate Program in Biophysical Sciences, The University of Chicago, Chicago, IL 60637, USA

2Department of Medicine, Biological Sciences Division, The University of Chicago, Chicago, IL 60637, USA

3Pritzker School of Molecular Engineering, The University of Chicago, Chicago, IL 60637, USA

4These authors contributed equally

5Technical contact

6Lead contact

*Correspondence: huangjun@uchicago.edu

**Correspondence: yfang1@medicine.bsd.uchicago.edu


Heterogeneous metabolism supports critical single-cell functions. Here, we describe deep-learning-enabled image analyses of a genetically encoded lactate-sensing probe which can accurately quantify metabolite levels and glycolytic rates at the single-cell level. Multiple strategies and test data have been included to obviate possible obstacles including successful sensor expression and accurate segmentation. This protocol reliably discriminates between metabolically diverse subpopulations which can be used to directly link metabolism to functional phenotypes by integrating spatiotemporal information, genetic or pharmacological perturbations, and real-time metabolic states.

Method diagram

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