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CryoSIM: super-resolution 3D structured illumination cryogenic fluorescence microscopy for correlated ultrastructural
Michael A Phillips1,2,3, Maria Harkiolaki3, David Miguel Susano Pinto1
1Micron Advanced Bio-imaging Unit, Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK.
Optica
|July 19, 2021
Summary
We developed cryoSIM, a super-resolution microscope for 3D cryo-fluorescence imaging. This instrument enables high-resolution visualization of cryopreserved specimens, correlating with ultrastructure for molecular detection.
Area of Science:
- Cellular and Molecular Imaging
- Biophysics
- Microscopy Instrumentation
Background:
- Rapid cryopreservation is crucial for preserving cellular structures.
- Existing cryo-fluorescence microscopes offer limited resolution.
- A need exists for higher resolution imaging correlated with ultrastructure.
Purpose of the Study:
- To develop a 3D super-resolution fluorescence cryo-imaging microscope (cryoSIM).
- To enable correlation of molecular localization with cellular ultrastructure.
- To democratize access to super-resolution cryo-imaging.
Main Methods:
- Construction of cryoSIM using off-the-shelf components.
- Development of open-source Python control software.
- Integration of super-resolution fluorescence imaging with cryo-preservation.
Main Results:
- Successful development of the cryoSIM instrument.
- Demonstration of 3D super-resolution fluorescence cryo-imaging.
- Provision of open-source instructions and software for replication.
Conclusions:
- cryoSIM bridges the resolution gap in cryo-fluorescence microscopy.
- The instrument facilitates correlation with cryo-electron microscopy and cryo-soft X-ray tomography.
- cryoSIM democratizes super-resolution imaging of cryopreserved biological specimens.
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