High speed biological imaging at and beyond the diffraction limit

Hari Shroff

National Institute of Biomedical Imaging and Bioengineering

National Institutes of Health

Bethesda, MD USA

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.

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. 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 diSPIM). Applications of this technology will be presented, including computational methods for untwisting worm embryos.