Title : Biological Imaging at High Spatiotemporal Resolution
Abstract : I will discuss our latest attempts to improve fluorescence microscopy techniques for following live biological phenomena at high spatiotemporal resolution. Emphasis will be given to structured illumination microscopy (SIM) and light-sheet microscopy (LSFM) techniques.
SIM1 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 thicker2-4 than possible with conventional SIM, as well as hardware modifications that enable effectively ‘instant’ SIM3,5 imaging at rates 10-100x faster than other SIM. New applications of instant SIM, including combination with total internal reflection (TIRF) and with adaptive optics6 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 nematodes7. I will discuss progress that quadruples the axial resolution of iSPIM by utilizing a second specimen view, thus enabling imaging with isotropic spatial resolution (dual-view iSPIM, or diSPIM8,9). Newer multiview results with more objectives10 and more views11, further improving spatial resolution, will also be shown. Applications of these technologies will be presented, including computational methods for untwisting worm embryos12, with the goal of building a neurodevelopmental atlas with subcellular resolution13.
Time permitting, I will also discuss 3 new and unpublished research projects in my lab: (1) rapid, sensor-less adaptive optics for use in SIM and LSFM; (2) 3D orientation sensing via multi-view fluorescence polarization; (3) structured illumination microcopy with ~5-fold improvement in axial resolution.