A key part of drug screening programs is monitoring effects of candidate drugs on mitochondrial health. Mitochondrial activity can be readily analyzed using fluorescent dyes available in a variety of colors. These are often used in conjunction with a nuclear counterstain, such as DAPI, which enables identification of all cells in the image. Once the cells have been identified via their nuclear stain in one fluorescence channel of an imaging system, their mitochondrial activity can be measured on a per-cell basis as fluorescence in a second channel corresponding to the specific mitochondrial dye used. However, nuclear counterstaining adds an extra series of cumbersome steps to the experimental workflow and delays results. StainFree™ Technology from Molecular Devices frees researchers from the inconvenience of nuclear counterstaining by identifying cells imaged with the transmitted light channel of the SpectraMax® MiniMax™ 300 Imaging Cytometer.
This application highlight shows how the SpectraMax® i3 Multi-Mode Detection Platform can be used to assess mitochondrial toxicity with cellular imaging. Valinomycin, an ionophore antibiotic, was used to disrupt mitochondrial function in PC12 cells, and MitoTracker Deep Red FM was used to monitor the effects. Analyses using StainFree and fluorescent nuclear dye to identify the cells were compared and results evaluated using SoftMax® Pro Software. The SpectraMax MiniMax 300 Imaging Cytometer and SoftMax Pro Software provide a complete solution for image acquisition and analysis for rapid quantitation of mitochondrial activity in response to potentially toxic compounds.
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