Mitochondria and mitochondrial function are highly sensitive indicators of cell health and stress. Depolarization of the inner mitochondrial membrane potential is a reliable indicator of mitochondrial dysfunction, which has been increasingly implicated in drug toxicity.

The HCS Mitochondrial Health Kit was developed for simultaneous quantitative measurements of mitotoxicity and cytotoxicity in the same cell. The MitoHealth stain (included in the kit) accumulates in the mitochondria of live cells in an amount that is proportional to the mitochondrial membrane potential. Other probes included in the kit include the Image-iT® DEAD Green™ viability stain, which enables discrimination of dead cells by plasma membrane integrity, and Hoechst 33342, which stains DNA in live and dead cells and is provided as a segmentation tool for automated image analysis. The staining pattern achieved using the dyes in this kit is retained after formaldehyde fixation and detergent permeabilization and is amenable to subsequent immunocytochemical detection of additional targets.

HCS Mitochondrial Health Kit

Figure 4—Dose response for valinomycin in HeLa cells using the HCS Mitochondrial Health Kit. HeLa cells were treated with valinomycin at final concentrations between 2 nM and 120 µM. Cells were incubated for 24 hours at 37°C/5% CO2 and assayed using the HCS Mitochondrial Health Kit. The Thermo Scientific Cellomics® ArrayScan® VTI platform with Compartmental Analysis BioApplication was used for analysis. Dose response curves generated by non-linear regression with GraphPad PRISM® were used to determine EC50 values for valinomycin-induced loss of plasma membrane integrity and mitochondrial membrane potential with increasing concentrations of valinomycin. Note: Since valinomycin is less soluble in DMSO at concentrations above 120 µM, data points beyond this concentration are not shown in the figure.
Product                               Cat. no.
Emission Color*
Ex/Em† Live‡ Fixed§ Fixable**
HCS Mitochondrial Health KitH10295       350/461
* Gray represents fluorescence emission that is beyond the limit of human vision. † Ex/Em = Fluorescence excitation and emission maxima in nm. ‡ Stains live cells. § Stains fixed cells. ** Staining pattern is fixable. NA = Not applicable.