Application Notes

Visualization of cell death pathways by live cell imaging: Analysis of apoptosis using mitochondrial membrane potential and annexin V

November 2025

The development of fluorescent probes and microscopy technology have made significant contributions to medicine and biology. They enable real-time observation of the movements of molecules and organelles within living cells, advancing our understanding of cell death pathways and drug mechanisms of action. Furthermore, advances in image processing and analysis technology have enabled label-free cell counting directly from brightfield images. This allows for the simultaneous acquisition of more information, such as cell morphology observation and cell counting, while reducing phototoxicity in live-cell imaging. This application note presents an example of analyzing early apoptosis with low phototoxicity by visualizing mitochondrial membrane potential (MMP) and annexin V using fluorescent probes, and measuring cell numbers using label-free Volume Contrast (VC) imaging. Furthermore, we analyzed differences in cellular response to three drugs: etoposide, staurosporine, and antimycin A.

Keywords: Early apoptosis, Mitochondrial membrane potential, Cell death pathway, Fluorescent probe, Live cell imaging, Volume contrast, Image analysis, Label-free cell counting