Fluorescence microscopy image analysis
automation - time series - physiology

TMRM FLIPR technique - mitochondrial assay

bis-oxonol and tetramethylrhodamine methyl ester (TMRM)

Mitochondrial membrane potential assay

Absolute and Unbiased Mitochondrial Membrane Potential Assay

Commonly used mitochondrial membrane potential (ΔψM) assays often lead to data misinterpretation. This is because all of the used probes are influenced by multiple properties of cells other than ΔψM.

Our technology is based on biophysical modeling of ΔψM probe behavior to back-calculate potentials from time courses of fluorescence intensities. This enables absolute millivolts readout, and an unbiased comparison of different samples, accounting for geometric, binding and ΔψP effects.

The Theory

Theory of the TMRM/FLIPR technique

How absolute millivolts are calculated

See how effects of ΔψP, cell and mitochondrial geometry and fluorescence background accounted for to calculate mV values.

In Practice

Membrane Potential Calibration Wizard

The Membrane Potential Calibration Wizard

This interactive dialog works hand in hand with the provided protocol to calibrate potentials without theoretical knowledge.

Try it out

See the technology in publications

Published in-depth protocol (61)
In human stem cells (14)
In rat neurons (17)
In human and rodent β-cells (26,31,40)
In osteoblasts (40)
In MCF7 (49)
In hepatocellular carcinoma (51)
In H29C, BJ1, HepG2 cells, primary cardiomyocytes (61)

Mitochondrial Membrane Potential Assay Protocol

Lerner and Gerencser Mitochondrial Membrane Potential

Protocol

Protocol explaining experimental design, microscopy setup and image analysis

The development of the unbiased, absolute mitochondrial membrane potential assay has been supported by:

SBIR/STTR

NIDA