Fluorescence Time-lapse Imaging of Entosis Using Tetramethylrhodamine Methyl Ester Staining

Emir Bozkurt; Heiko Düssmann; Jochen H M Prehn

doi:10.21769/BioProtoc.4564

Fluorescence Time-lapse Imaging of Entosis Using Tetramethylrhodamine Methyl Ester Staining

Bio Protoc. 2022 Dec 5;12(23):e4564. doi: 10.21769/BioProtoc.4564.

Authors

Emir Bozkurt¹, Heiko Düssmann¹, Jochen H M Prehn¹

Affiliation

¹ Department of Physiology and Medical Physics, Centre for Systems Medicine, Royal College of Surgeons in Ireland, Dublin, Ireland.

Abstract

Entosis is a process where a living cell launches an invasion into another living cell's cytoplasm. These inner cells can survive inside outer cells for a long period of time, can undergo cell division, or can be released. However, the fate of most inner cells is lysosomal degradation by entotic cell death. Entosis can be detected by imaging a combination of membrane, cytoplasmic, nuclear, and lysosomal staining in the cells. Here, we provide a protocol for detecting entosis events and measuring the kinetics of entotic cell death by time-lapse imaging using tetramethylrhodamine methyl ester (TMRM) staining. This protocol was validated in: J Cell Biol (2021), DOI: 10.1083/jcb.202010030.

Keywords: Confocal fluorescence microscopy; Entosis; Entotic cell death; TMRM; Time-lapse imaging; Venus.