PROJECT SUMMARY – ADVANCED BIOPHOTONICS The Advanced Biophotonics Shared Resource (BPHO) provides imaging and analysis instrumentation coupled with technical and collaborative support staff for advanced light and electron microscopy of biological systems. In addition to the Director, the BPHO is staffed by three full-time imaging specialists who have expertise in electron and light microscopy, as well as data analysis. Salk Cancer Center (SCC) members use the facility for high-throughput imaging assays, high-resolution imaging of live cell and tissue dynamics, super-resolution microscopy, large 3D volume imaging of tissues, electron microscopy analysis of subcellular morphology and protein distribution, and automated computational image processing, visualization, and analysis. The BPHO is also actively pursuing and developing new cutting-edge imaging and analysis methodologies to better serve the needs of SCC researchers, such as cryo-correlative light and electron microscopy, light-sheet imaging of cleared and expanded tissues, and machine-learning based processing, segmentation, and analysis of light and electron microscope images. BPHO is committed to providing SCC members: 1) Access to light and electron microscopes, specialized sample preparation reagents and technologies, and computational hardware and software for analysis and visualization 2) Free one-on-one training on all microscopes, as well as image processing and analysis software 3) Consulting and collaborative support for experimental design and implementation of imaging and analysis experiments 4) Sample preparation for electron microscopy, tissue clearing, and expansion microscopy 5) Workshops and demos with advanced microscopy and software technologies 6) Weekly open-door imaging boot camps on advanced imaging and image processing techniques 7) A monthly Biophotonics scientific seminar series followed by town-hall discussion with BPHO staff and users Utilization of the imaging technologies and expertise in this Shared Resource is high. During the current funding period, eighty-one percent (81%) of the SCC members have benefited from utilization of the facility. This accounts for approximately 61% of total BPHO utilization.