Super-Resolution Microscopy

Super-resolution microscopy is an optical imaging technology that overcomes the diffraction limit of light and allows the visualization of subcellular structures and dynamics in greater detail than can be achieved with conventional optical microscopy. A resolution of 30 nm is possible using STED (stimulated emission depletion) with nanoscopy. A nanoscale resolution reveals information about subcellular structures and interactions with unprecedented detail. Super-resolution microscopy enables groundbreaking discoveries in the fields of virology, immunology, neuroscience, and cancer research.

For more information about super-resolution microscopy, refer to the Science Lab articles mentioned above or go to: The Microscopy Knowledge Portal

Super-resolution microscopy can be performed in several ways. Here are the 3 more common methods offered by Leica Microsystems:

1. The LIGHTNING detection concept enables super-resolution confocal microscopy. Unlike traditional deconvolution techniques that use a global set of parameters for the full image, it is based on an adaptive deconvolution process which uses a voxel-specific decision mask for finding the optimal deconvolution parameters . Thanks to powerful GPU computing, LIGHTNING works in near real-time for multiple colors simultaneously. In combination with a resonant scanner, it also provides a large field of view and high frame rate. For more information about LIGHTNING, refer to the following: Obtain maximum information from your specimen with LIGHTNING & Image information extraction by adaptive deconvolution
2. The STED method along with confocal also allows super-resolution fluorescence microscopy. STED works with multiple channels and approaches isotropic super-resolution imaging in three dimensions. STED is based on the illumination of a diffraction-limited spot with a fluorophore-exciting wavelength and simultaneous illumination of a ring-shaped area with wavelengths that cause de-excitation or stimulated emission. This arrangement renders only a sub-diffraction area in the excited state and consequently allows probing with higher resolution than that of typical diffraction-limited optical microscopy. The unique TauSTED functionality from Leica Microsystems even allows for the increased STED resolution while eliminating undesired background noise .

For more information about STED, refer to: Video Talk on Super-Resolution: Overview and Stimulated Emission Depletion (STED) Microscopy & TauSTED: pushing STED beyond its limits with lifetime

Whereas super-resolution microscopy exploits visible light optics to visualize a sample, electron microscopy exploits a beam of electrons for imaging a sample. Both techniques have their pros and cons, yet only super-resolution microscopy offers the advantages of experimenting with living specimens, labeling multiple molecular targets to give them specific contrast, and convenient sample preparation.