Ultra-fast GSDIM super resolution microscopy using a SPAD-array camera (GSDIM)

The ability to directly visualize nanoscopic cellular structures and their spatial relationship in fixed specimens, and to combine this with high resolution imaging of molecular action and interaction in living cells, will increase the understanding of molecular processes in living cells.

Recently, several new technologies for super-resolution have been developed. Stochastic optical reconstruction microscopy, STORM and the similar photoactivated localization microscopy, PALM have been developed to quantitatively probe cellular structures and their interactions. To facilitate STORM/PALM imaging, photoswitchable probes were generated in several distinct colors. With this technology, whole-cell images were obtained with a spatial resolution of 20-30 nm and 60-70 nm in the lateral and axial dimensions, respectively.

An alternative approach, ground state depletion followed by individual molecule return, GSDIM, offers the important advantage that it allows the use of nonswitchable standard fluorophores.