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Updated: Jun 14, 2026

Simultaneous Measurement of Mitochondrial Calcium and Mitochondrial Membrane Potential in Live Cells by Fluorescent Microscopy
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Imaging mitochondrial membrane potential via concentration-dependent fluorescence lifetime changes.

Dilizhatai Saimi1, Luc Reymond2, Tursunjan Aziz3

  • 1College of Future Technology, Institute of Molecular Medicine, National Biomedical Imaging Center, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Peking University, Beijing, China.

Nature Communications
|December 12, 2025
PubMed
Summary
This summary is machine-generated.

Researchers developed PK Mito Deep Red (PKMDR), a novel mitochondrial probe. Its fluorescence lifetime accurately measures mitochondrial membrane potential (Δψm), revealing metabolic heterogeneity in cells and tissues.

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Biophysics

Background:

  • Mitochondria are vital for cellular metabolism and energy production.
  • Existing fluorescence tools for mitochondrial imaging have limitations in spatial-temporal resolution.
  • Direct reporters for mitochondrial status, particularly membrane potential, are needed.

Purpose of the Study:

  • To introduce PK Mito Deep Red (PKMDR), a novel fluorescent probe for mitochondrial imaging.
  • To establish PKMDR's fluorescence lifetime as a sensitive indicator of mitochondrial membrane potential (Δψm).
  • To demonstrate the utility of PKMDR-based fluorescence lifetime imaging microscopy (FLIM) for visualizing metabolic heterogeneity.

Main Methods:

  • Development and characterization of PKMDR, a low-phototoxicity mitochondrial probe.
  • Utilizing fluorescence lifetime imaging microscopy (FLIM) to measure Δψm.
  • Application of PKMDR-FLIM in live cells, organoids, and tissues for time-lapse imaging.

Main Results:

  • PKMDR fluorescence lifetime is a sensitive indicator of Δψm, correlating with mitochondrial respiration and oxidative phosphorylation.
  • PKMDR-FLIM effectively visualizes heterogeneous Δψm across individual cells, organoids, and tissues.
  • The method reveals metabolic differences between peripheral and perinuclear mitochondria.

Conclusions:

  • PKMDR is a robust fluorescent probe for assessing mitochondrial membrane potential.
  • PKMDR-FLIM provides high spatiotemporal resolution for visualizing mitochondrial metabolic status.
  • This technique offers new insights into cellular and tissue metabolic heterogeneity.