Optical estimation of absolute membrane potential using fluorescence lifetime imaging.

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ID: 55854
2019
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Abstract
All cells maintain ionic gradients across their plasma membranes, producing transmembrane potentials (V). Mounting evidence suggests a relationship between resting V and the physiology of non-excitable cells with implications in diverse areas, including cancer, cellular differentiation, and body patterning. A lack of non-invasive methods to record absolute V limits our understanding of this fundamental signal. To address this need, we developed a fluorescence lifetime-based approach (VF-FLIM) to visualize and optically quantify V with single-cell resolution in mammalian cell culture. Using VF-FLIM, we report V distributions over thousands of cells, a 100-fold improvement relative to electrophysiological approaches. In human carcinoma cells, we visualize the voltage response to growth factor stimulation, stably recording a 10-15 mV hyperpolarization over minutes. Using pharmacological inhibitors, we identify the source of the hyperpolarization as the Ca-activated K channel K3.1. The ability to optically quantify absolute V with cellular resolution will allow a re-examination of its signaling roles.
Reference Key
lazzaridean2019opticalelife Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Lazzari-Dean, Julia Rose;Gest, Anneliese M M;Miller, Evan W;
Journal eLife
Year 2019
DOI
10.7554/eLife.44522
URL
Keywords
physics of living systems biochemistry human chemical biology

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