White Paper: Materials

Single-molecule Localisation Microscopy (Smlm): How to Overcome the Abbe Diffraction Limit?


The Abbe diffraction barrier, which fundamentally restricted the lateral optical resolution to 200 – 300 nm until the early 1990s, can be overcome by super-resolution optical fluorescence imaging techniques. These techniques have been so groundbreaking that the Nobel Prize 2014 in Chemistry was awarded to Eric Betzig, Stefan Hell and W. E. Moerner “for the development of super-resolved fluorescence microscopy”.

There are two different techniques commonly used so far; one is based on point spread function engineering, the other on single-molecule localization. Stefan Hell’s stimulated emission depletion (STED) microscopy belongs to the first class and is a so called deterministic super-resolution technique. In contrast, Eric Betzig’s photoactivated localization microscopy (PALM) is a stochastic technique. This whitepaper looks at the technique of single-molecule localisation microscopy, how it works and the benefits it can bring you.

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