Measuring abscission spatiotemporal dynamics using quantitative high-resolution microscopy

Abstract

The spatiotemporal characteristics of ESCRT (Endosomal Sorting Complex Required for Transport)-mediated mammalian cytokinetic abscission have been studied in recent years using quantitative high-resolution light microscopy techniques. Here we describe how to apply spinning disk live cell imaging and structured illumination microscopy (SIM) to define the dynamics and structural organization of abscission and of proteins involved in abscission in a quantitative manner. We further provide a protocol to correlate the structural data, obtained by SIM, to the dynamic information obtained by live cell recordings.

Keywords: Cell cycle; Cell division; Cytokinesis; ESCRT; Intercellular bridge; Live cell imaging; Membrane fission; Structured illumination microscopy.