Smart lattice light-sheet microscopy for imaging rare and complex cellular events

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Abstract
Light-sheet microscopes enable rapid high-resolution imaging of biological specimens; however, biological processes span spatiotemporal scales. Moreover, long-term phenotypes are often instigated by rare or fleeting biological events that are difficult to capture with a single imaging modality. Here, to overcome this limitation, we present smartLLSM, a microscope that incorporates artificial intelligence-based instrument control to autonomously switch between epifluorescent inverted imaging and lattice light-sheet microscopy (LLSM). We apply this approach to two unique processes: cell division and immune synapse formation. In each context, smartLLSM provides population-level statistics across thousands of cells and autonomously captures multicolor three-dimensional datasets or four-dimensional time-lapse movies of rare events at rates that dramatically exceed human capabilities. From this, we quantify the effects of Taxol dose on spindle structure and kinetochore dynamics in dividing cells and of antigen strength on cytotoxic T lymphocyte engagement and lytic granule polarization at the immune synapse. Overall, smartLLSM efficiently detects rare events within heterogeneous cell populations and records these processes with high spatiotemporal four-dimensional imaging over statistically significant replicates.
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Funding Information
  • Kinship Foundation (Searle Scholars Program)
  • Arnold and Mabel Beckman Foundation (Beckman Young Investigator, Beckman Young Investigator Award)
  • David and Lucile Packard Foundation (Packard Fellowship in Science and Engineering)
  • U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (1DP2GM136653-01)
  • Howard Hughes Medical Institute

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