Since Robert Hooke’s first description of a cell in Micrographia 350 years ago, microscopy has played an important role in understanding the rules of life. Far-field fluorescence microscopy is a powerful approach in biological and biomedical research due to its live cell compatibility and molecular specificity. A major hurdle over the last century has been the limited resolution due to the diffraction of light. Super-resolution microscopy methods, such as single-molecule localization microscopy, overcome this fundamental barrier to improve the resolution limit (250-600 nm, lateral-axial) down to a few nanometers. The development and application of SMLM majorly focus on fixed cells in thin samples and cellular structures that lie on/close to the coverslip surface and therefore, the profound impact of SMLM on biophysical, medical, and biological research has yet to fully unfold.
I will discuss some of our most recent projects which explore newly available sensors/devices such as high-speed sensors and deformable mirrors, analytical methods such as deep learning, and novel optical instrumentation to allow SMLM imaging in live cells and tissue specimens. I will show the capabilities of these new imaging systems in revealing the fine details of subcellular structures from a diverse set of biological systems including viruses, bacteria, yeasts, mammalian cells, and brain sections.