Objective: to develop a method which measures mitochondrial membrane potential of a group of cells.
Feature 1: measure the fluorescence of cationic probe in extracellular medium instead of the fluorescence of cationic probe in mitochondria which is likely to undergo self-quenching. Feature 2: consider the effect of plasma membrane potential, efflux pump on fluorescence distribution. Feature 3: measure mitochondrial membrane potential for a group of intact cells instead of isolated mitochondria or single cell. |
Procedure:
1. Design a set of experiments which measure extracellular probe concentrations where mitochondrial membrane potential plays a role on the distribution of probe 2. Establish a mathematical model to describe the experimental system 3. Estimate mitochondrial membrane potential by fitting the model to experimental data 4. Validate the model by predicting experimental results using the estimated values. Model: A model based on nernst equation and fick's law was established to describe the temporal distrubition of cationic probe in extracellular medium. Experiments: A set of experiments were designed to monitor the temporal distribution of rhodamine probes in extracellular medium in the prensents of cells. |
Application
The Estimation of Mitochondrial Membrane Potential
in Cultured Bovine Pulmonary Arterial Endothelial Cells
Related Publication:
Quantifying mitochondrial and plasma membrane potentials in intact pulmonary arterial endothelial cells based on extracellular disposition of rhodamine dyes
Gan Z, Audi SH, Bongard RD, Gauthier KM, Merker MP.
Am J Physiol Lung Cell Mol Physiol. 2011 May;300(5):L762-72
Quantifying mitochondrial and plasma membrane potentials in intact pulmonary arterial endothelial cells based on extracellular disposition of rhodamine dyes
Gan Z, Audi SH, Bongard RD, Gauthier KM, Merker MP.
Am J Physiol Lung Cell Mol Physiol. 2011 May;300(5):L762-72