CryoSIM: super-resolution 3D structured illumination cryogenic fluorescence microscopy for correlated ultrastructural imaging

• DOI: 10.1364/optica.393203
• Publication Date: 2020-06-08T16:30:10Z
• Authors: Michael A. Phillips; Maria Harkiolaki; David Miguel Susa...; Richard M. Parton; Ana Palanca; Manuel Garcia-Moreno; Ilias Kounatidis; John W. Sedat; David I. Stuart; Alfredo Castello; Martin J. Booth; Ilan Davis; Ian M. Dobbie
• Keywords: 0301 basic medicine; 570; Communications Technologies; 0303 health sciences; 1.1 Normal biological development and functioning; 610; Bioengineering; Optical Physics; 1.5 Resources and infrastructure (underpinning); Article; 03 medical and health sciences; Underpinning research; Generic health relevance; Electrical and Electronic Engineering

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.