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

DOI: 10.1364/OPTICA.393203 DOI Help

Authors: Michael A. Phillips (University of Oxford; Wellcome Centre for Human Genetics; Diamond Light Source) , Maria Harkiolaki (Diamond Light Source) , David Miguel Susano Pinto (University of Oxford) , Richard M. Parton (University of Oxford) , Ana Palanca (Universidad de Cantabria; University of Oxford) , Manuel Garcia-Moreno (University of Oxford) , Ilias Kounatidis (Diamond Light Source) , John W. Sedat (University of California, San Francisco) , David I. Stuart (Wellcome Centre for Human Genetics; Diamond Light Source) , Alfredo Castello (University of Oxford) , Martin J. Booth (University of Oxford) , Ilan Davis (University of Oxford) , Ian Dobbie (University of Oxford)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Optica , VOL 7

State: Published (Approved)
Published: July 2020
Diamond Proposal Number(s): 18319 , 18757

Open Access Open Access

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.

Subject Areas: Technique Development, Biology and Bio-materials


Instruments: B24-Cryo Soft X-ray Tomography

Added On: 15/04/2021 11:35

Documents:
optica-7-7-802.pdf

Discipline Tags:

Technique Development - Life Sciences & Biotech Life Sciences & Biotech

Technical Tags:

Imaging Microscopy Tomography Cryo Electron Tomography (Cryo ET) Electron Microscopy (EM) Cryo Electron Microscopy (Cryo EM)