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Building a super-resolution fluorescence cryomicroscope.

Mart G F Last¹, Lenard M Voortman¹, Thomas H Sharp²

¹Department of Cell and Chemical Biology, Leiden University Medical Centre, Leiden, The Netherlands.

Methods in Cell Biology

|May 5, 2024

Summary

This summary is machine-generated.

We developed the "cryoscope," a novel microscope for super-resolution fluorescence microscopy on cryo-electron microscopy samples. This innovation allows high-resolution imaging of biological structures with precise labeling within established workflows.

Keywords:

Correlative light and electron microscopy Microscope construction Super-resolution cryoCLEM

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Area of Science:

Biophysics
Cell Biology
Microscopy

Background:

Correlated super-resolution fluorescence microscopy (SMLM) and cryo-electron microscopy (cryo-EM) offer high labeling specificity and resolution.
Integrating these techniques requires specialized hardware capable of imaging cryogenic samples with super-resolution fluorescence.

Purpose of the Study:

To design and describe a novel microscope, the "cryoscope," for SMLM of cryo-EM samples.
To integrate SMLM into existing cryo-EM workflows.

Main Methods:

Design and construction of the "cryoscope" microscope.
Application of SMLM for imaging fluorescently labeled vimentin in U2OS cells on EM grids.
3D modeling of the microscope's complete design.

Main Results:

Demonstration of the cryoscope's capability for high-resolution imaging of biological samples.
Successful imaging of fluorescently labeled vimentin within U2OS cells.
Provision of detailed 3D models for the microscope's design.

Conclusions:

The cryoscope enables correlated SMLM and cryo-EM imaging by combining high labeling specificity and resolution.
The developed microscope integrates seamlessly into established cryo-EM workflows.
Detailed design information is provided for the cryoscope.