Microscopy technique enables 3D super-resolution nanometre-scale imaging

To show that 3D imaging with MIET-SMLM is compatible with biological samples, ce

To show that 3D imaging with MIET-SMLM is compatible with biological samples, cells were seeded on a cover glass coated with 10 nm of gold and 5 nm of SiO2 using standard immunofluorescence sample prepa-ration procedure. The artistic rendering illustrates cells imaging on a gold surface resolving microtubules network and clathrin coated pits. Photo: Alexey Chizhik

Research team led by Göttingen University combine two techniques to achieve isotropic super -resolution imaging 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. The results were published in Science Advances.

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 Dr Jan Christoph Thiele, first author of the publication, Göttingen University. -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 Dr Oleksii Nevskyi, one of the corresponding authors.

-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 Professor 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.

Original publication: Thiele et al, Isotropic three-dimensional dual-color super-resolution microscopy with metal-induced energy transfer, Science Advances 2022. DOI: 10.1126/sciadv.abo2506

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