Mapping the inner world of the cell
Seminar | September 12 | 12-1 p.m. | 106 Stanley Hall
Bo Huang, University of Califronia, San Francisco
Cellular processes are orchestrated by a large number of biomolecules in a spatially and temporally coordinated manner within a tiny volume. To uncover the underlying organizational principles and their functional relevance, we take microscopy visualization as the primary approach to systematically map the spatial localization, temporal dynamics and activity profiles of proteins and nucleic acids. For this purpose, we have developed a fragment-tagging approach which fluorescently labels target proteins using our engineered split fluorescent proteins. This approach has enabled systematic generation of knock-in cell lines with endogenously labeled proteins through CRISPR/Cas9-medied gene editing. To perform high-throughput live imaging of such cell lines, we have developed an epi-illumination selective-plane illumination microscopy (eSPIM) system. Based on the optical frame of a conventional epi-fluorescence microscope, eSPIM is capable of producing high-resolution volumetric images of cells and organoids in conventional multiwell plates. Taken together, these techniques build up a toolbox for the drawing of cellular and subcellular maps.