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. 2021 Jun 30;9:692776. doi: 10.3389/fcell.2021.692776

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Copyright © 2021 Aklima, Onojima, Kimura, Umiuchi, Shibata, Kuraoka, Oie, Suganuma and Ohta.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Time course of changes in Δψ_m in isolated mitochondria. (A,B) Optical images of isolated mitochondria adsorbed on a cover slip. (A) Brightfield and (B) fluorescence images of the same microscopic field as (A). Bar, 3 μm. (C) Time-resolved fluorescence images of TMRE in a single mitochondrion. TMRE fluorescence was monitored in the presence of malate (5 mM). The arrows show the onset of depolarization and polarization. The time interval between images is 3 s. Bar, 2 μm. (D) Membrane potential (Δψ_m) in a single mitochondrion in response to malate addition. The vertical axis represents the fluorescence intensity of TMRE in a single mitochondrion, normalized by TMRE fluorescence in the buffer (TMRE Ratio). At t = 2 min, 5 mM malate was added. The fluctuations of Δψ_m significantly depend on individual mitochondria. Δψ_m in mitochondria 1–3 is fluctuating, but not in mitochondrion 4.