Pushing the envelope in biological microscopy: High speed imaging at and beyond the diffraction limit

Director's Seminar Series Dr. Hari Shroff will discuss his lab's, the Section of High Resolution Optical Imaging (HROS), 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. Dr. Shroff will present SIM implementations that enable resolution doubling in live samples> 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. New 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 inverted selective plane illumination microscopy (iSPIM), and subsequent application to the noninvasive study of neurodevelopment in nematodes. Dr. Shroff will discuss progress that quadruples the axial resolution of iSPIM by using a second specimen view, enabling imaging with isotropic spatial resolution (dual-view iSPIM, or diSPIM). Newer multiview microscopes with more objectives and more views, further improving spatial resolution, will also be described. Applications of these technologies will be presented, including computational methods for untwisting worm embryos and calcium imaging in freely moving embryos. Finally, Dr. Shroff will discuss the cur...
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