Abstract

A Revolutionary Miniaturised Instrument For Single-Molecule Localization Microscopy And FRET

Achillefs Kapanidis, Professor, University of Oxford

The ability to detect single fluorescent molecules has advanced substantially our understanding of biological mechanisms and structures, and has enabled localization microscopies that bypass the diffraction limit to produce stunning images with resolution as high as 10nm. However, the instruments currently available for single-molecule fluorescence detection and super-resolution imaging are large, complex, and require specialized infrastructure that invariably confines them in user facilities or specialized rooms. Here we describe the development of the first desktop single-molecule fluorescence imager for super-resolution imaging and single-molecule FRET applications. The main microscope module is compact (just 21x21x15cm) and features several means of stabilization against vibrations, temperature-dependent drift, and focal drift, allowing stable operation on a regular laboratory bench for hours. The large field-of-view of the microscope allows localization-based super-resolution imaging of large cells, as well as high-throughput real-time analysis of conformational dynamics and biochemical reactions using single-molecule FRET. We anticipate that our compact microscope will popularize the use of single-molecule methods and super-resolution imaging in many research communities.