Semi-automated Detection of Late Cytokinetic Bridges

During cytoplasmic division, the intercellular bridge is formed with a central midbody. In our opinion, more tools are required for investigation of the late molecular events leading to cytokinetic abscission. In this chapter, we briefly review the many available methods to screen for cytokinetic defects and present a protocol for image-based detection of late cytokinetic bridges. A simple immunofluorescence staining of the fixed cytoplasms, nuclei, and the midbodies, analyzed with tailored software, can reproducibly estimate the frequency of intercellular bridges. Here, we describe data collection and analysis in single cells upon inhibition by RNA interference using fluorescence microscopy.