Isotropic three-dimensional dual-color super-resolution microscopy with metal-induced energy transfer
Jan Christoph Thiele, Marvin Jungblut, Dominic A. Helmerich, Roman Tsukanov, Anna Chizhik, Alexey I. Chizhik, Martin J. Schnermann, Markus Sauer, Oleksii Nevskyi, Jörg Enderlein
Abstract
Over the past two decades, super-resolution microscopy has seen a tremendous development in speed and resolution, but for most of its methods, there exists a remarkable gap between lateral and axial resolution, which is by a factor of 2 to 3 worse. One recently developed method to close this gap is metal-induced energy transfer (MIET) imaging, which achieves an axial resolution down to nanometers. It exploits the distance-dependent quenching of fluorescence when a fluorescent molecule is brought close to a metal surface. In the present manuscript, we combine the extreme axial resolution of MIET imaging with the extraordinary lateral resolution of single-molecule localization microscopy, in particular with direct stochastic optical reconstruction microscopy ( d STORM). This combination allows us to achieve isotropic three-dimensional super-resolution imaging of subcellular structures. Moreover, we used spectral demixing for implementing dual-color MIET- d STORM that allows us to image and colocalize, in three dimensions, two different cellular structures simultaneously.