Multiscale Biological Imaging at High Spatiotemporal Resolution

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Structured illumination microscopy, light sheet microscopy, adaptive optics, neurodevelopment, developmental biology.

Online Seminar with Hari Shroff, from National Institute of Biomedical Imaging and Bioengineering National Institutes of Health Bethesda

Imaging biology at multiple spatial and temporal scales demands innovation in imaging technology. For the last ten years, our lab has focused on developing better microscopy techniques that span the subcellular to organismal spatial scale, and the millisecond to multi-hour temporal scale.

I will discuss our efforts to improve structured illumination microscopy (SIM) and light-sheet microscopy. SIM doubles the spatial resolution of light microscopy, requiring lower light intensities and acquisition times than other super-resolution techniques. I will present SIM implementations that enable resolution doubling in live volumes > 10-20x thicker than possible with conventional SIM, as well as hardware modifications that enable effectively ‘instant’ SIM imaging at rates 10-100x faster than other SIM. Newer applications of instant SIM, including combination with total internal reflection (TIRF) and with adaptive optics will also be discussed.

The second half of the talk will focus on the development of dual-view inverted selective plane illumination microscopy (diSPIM), and subsequent application to the noninvasive study of neurodevelopment in nematodes. Newer multiview results with more objectives and more views, further improving spatial resolution, will also be shown. Applications of these technologies will be presented, including computational methods for untwisting worm embryos, and calcium and behavioral imaging of moving embryos. Finally, recently developed methods for rapidly processing very large, TB-scale light-sheet datasets will be presented.

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