CryoSIM: super-resolution 3D structured illumination cryogenic fluorescence microscopy for correlated ultrastructural imaging
Michael A. Phillips, Maria Harkiolaki, David Miguel Susano Pinto, Richard M. Parton, Ana Palanca, Manuel García-Moreno, Ilias Kounatidis, John W. Sedat, David I. Stuart, Alfredo Castelló, Martin J. Booth, Ilan Davis, Ian M. Dobbie
Abstract
Rapid cryopreservation of biological specimens is the gold standard for visualizing cellular structures in their true structural context. However, current commercial cryo-fluorescence microscopes are limited to low resolutions. To fill this gap, we have developed cryoSIM, a microscope for 3D super-resolution fluorescence cryo-imaging for correlation with cryo-electron microscopy or cryo-soft X-ray tomography. We provide the full instructions for replicating the instrument mostly from off-the-shelf components and accessible, user-friendly, open-source Python control software. Therefore, cryoSIM democratizes the ability to detect molecules using super-resolution fluorescence imaging of cryopreserved specimens for correlation with their cellular ultrastructure.