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Journal of Biomedical Optics • Open Access

Real-time monitoring of changes in plasma membrane potential via imaging of fluorescence resonance energy transfer at individual cell resolution in suspension
Author(s): Tzachi Sabati; Bat-Sheva Galmidi; Alon Korngreen; Naomi Zurgil; Mordechai Deutsch

Paper Abstract

A method for monitoring heterogeneity in changes of plasma membrane potential (PMP) at an individual cell resolution while in suspension, utilizing a simple and low-cost wide-field illumination arrangement, is presented. The method is modeled via HEK-293 cell line in suspension, double stained with coumarin and oxonol (donor and acceptor), which were loaded into an array of nanoliter wells, each designed to preserve the individuality of the nontethered cell it holds during vigorous biomanipulation. Depolarization of PMP was induced by high K + solution, reducing the proximity between the membrane fluorophores and subsequently reducing the efficiency (E% ) of resonance energy transfer between them. Spatial plots of E% were produced from both images of fluorescence intensity and polarization. The spatial resolution of E% plots seem to be higher, and their contrast greater, when calculated from the polarization, rather than from the intensity of the fluorescence.

Paper Details

Date Published: 16 December 2013
PDF: 11 pages
J. Biomed. Opt. 18(12) 126010 doi: 10.1117/1.JBO.18.12.126010
Published in: Journal of Biomedical Optics Volume 18, Issue 12
Show Author Affiliations
Tzachi Sabati, Bar-Ilan Univ. (Israel)
Bat-Sheva Galmidi, Bar-Ilan Univ. (Israel)
Alon Korngreen, Bar-Ilan Univ. (Israel)
Naomi Zurgil, Bar-Ilan Univ. (Israel)
Mordechai Deutsch, Bar-Ilan Univ. (Israel)

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