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Cell BiologybioRxiv

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

M. A. Phillips, M. Harkiolaki, D. M. S. Pinto, R. M. Parton, A. Palanca, M. Garcia-Moreno, I. Kounatidis, J. Sedat, D. I. Stuart, A. Castello, M. J. Booth, I. Davis, I. M. Dobbie

Abstract

Rapid cryo-preservation of biological specimens is the gold standard for visualising 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 democratises the ability to detect molecules using super-resolution fluorescence imaging of cryo-preserved specimens for correlation with their cellular ultrastructure.

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