Over the last two decades, microscopy has seen unpre­cedented advances in speed and resolution. However, cellular structures are essentially three-dimen­sional, and conventional super-resolution techniques often lack the necessary resolution in all three directions to capture details at a nanometer scale. A research team led by Göttingen University, including the University of Würzburg and the Center for Cancer Research in the US, investi­gated a super-resolution imaging technique that involves combining the advantages of two different methods to achieve the same resolution in all three dimensions; this is isotropic resolution.

Despite tremendous improve­ments in microscopy, there still exists a remarkable gap between resolution in all three dimensions. One of the methods that can close this gap and achieves a resolution in the nanometer range is metal-induced energy transfer (MIET) imaging. The exceptional depth resolution of MIET imaging was combined with the extra­ordinary lateral reso­lution of single-molecule locali­zation microscopy, in particular with a method called direct stochastic optical recon­struction microscopy (dSTORM). The novel technique based on this combi­nation allows researchers to achieve isotropic three-dimen­sional super-resolution imaging of sub-cellular structures. In addition, the researchers implement dual-color MIET-dSTORM enabling them to image two different cellular structures in three dimensions, for example micro­tubules and clathrin coated pits – tiny structures within cells – that exist together in the same area.

“By combining the established concepts, we developed a new technique for super-reso­lution microscopy. Its main advantage is it enables extremely high resolution in three dimensions, despite using a relatively simple setup,” says Jan Christoph Thiele. “This will be a powerful tool with numerous appli­cations to resolve protein complexes and small organelles with sub-nano­meter accuracy. Everyone who has access to confocal microscope technology with a fast laser scanner and fluorescence lifetime measurements capa­bilities should try this technique,” says Oleksii Nevskyi.

“The beauty of the technique is its simpli­city. This means that researchers around the world will be able to implement the technology into their micro­scopes quickly,” adds Jörg Enderlein who led the research team at the Biophysics Institute, Göttingen University. This method shows promise to become a powerful tool for multiplexed 3D super-resolution micro­scopy with extra­ordinary high resolution and a variety of appli­cations in structural biology. (Source: GAU)

Reference: J. C. Thiele et al.: Isotropic three-dimensional dual-color super-resolution microscopy with metal-induced energy transfer, Sci. Adv. 8, abo2506 (2022); DOI: 10.1126/sciadv.abo2506

Link: Single Molecule Spectroscopy and Imaging for Biophysics and Complex Systems, III. Physical Institute, Göttingen University, Göttingen, Germany