Cellular function of ESCRTs
At the end of mammalian cell division, the two nascent daughter cells are connected by a narrow membrane tube called the intercellular bridge. Cutting of the intercellular bridge, termed abscission, is driven by the ESCRT complex. The spatiotemporal characteristics of abscission are ideal for high-end microscopy techniques. To elucidate the mechanism of ESCRTs in physiological context we therefore apply high-resolution microscopy techniques to visualize ESCRTs in mammalian cell abscission. To obtain an inclusive understanding of ESCRT function in abscission, we visualize abscission in both mammalian tissue culture cells and in a developmental system (zebrafish embryogenesis). By combining information obtained from a variety of microscopy tools including live cell imaging, X-ray tomography and Super resolution microscopy we generate mechanistic models for ESCRT-mediated membrane abscission.
Advancing microscopy
High-resolution fluorescence imaging, combined with labeling techniques available today have yet been able to provide the spatiotemporal resolution needed to document cellular processes at physiological conditions. Substituting the use of fluorescent protein tags (such as GFP) typically used in live cell applications with fluorescent dyes will improve the currently advanced imaging techniques considerably. The superb photophysical characteristics of Fl-dyes and their small size sizes make them superior probes for quantitative high-resolution fluorescence imaging, particularly in the study of macromolecular assemblies.
Our tool box
We invest a great effort in developing ways to resolve the structural organization of ESCRT complex in abscission at sufficient spatiotemporal resolution. Over the past few years we established a microscopy-based toolbox to reach our goal.
