Capturing Full-color in Super-resolution Microscopy

The nanoscale architecture of biological systems underlies and regulates essentially all molecular machinery. Every cellular structure has a complex nanoscale organization ranging from individual macromolecules that are a few nanometers in size (e.g. protein, DNA) to macromolecular assemblies that may span tens of nanometers. The major scientific challenge is to understand these macromolecular assemblages, their function and interactions in non-perturbed, structurally and dynamically complex cellular systems. The importance of nanoscopic imaging is underscored by the 2014 Nobel Prize in Chemistry for the invention of superresolution optical microscopy. However, the potential of this technology can be immensely enhanced. In this talk, I will introduce spectroscopic single molecular localization microscopy (sSMLM) that captures the full emission spectra of individual molecules while simultaneously localizing their spatial locations at a precision greatly exceeding the optical diffraction limit. To achieve this, sSMLM uses a dispersive optical component to separate the emitted photons into dedicated spatial and spectral imaging channels for simultaneous acquisition. Exploiting its unique advantages in capturing the full emission spectra of individual single molecular emissions, we demonstrate the utilization of sSMLM in (1) multi-color super-resolution imaging, (2) a new strategy towards three-dimensional (3D) super-resolution imaging, and final (3) significantly improved imaging resultion beyond 10 nm.