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Building a super-resolution fluorescence cryomicroscope

Last, M. G. F.; Voortman, L. M.; Sharp, T. H.

2023-11-21 molecular biology
10.1101/2023.11.21.567712 bioRxiv
Show abstract

Correlating super-resolution fluorescence microscopy with cryo-electron tomography is a recent advancement in the field of cryo-electron microscopy that enables targeted, high-resolution imaging of specific biomolecules of interest. Critical to this approach is that the cryo-correlated light and electron microscopy (cryoCLEM) workflow requires samples to be cryogenically fixed prior to imaging, and thus a fluorescence microscope is required that can maintain the cryogenically preserved state of the sample while also being capable of super-resolution imaging. In this report, we outline the blueprint of a microscope that was designed for single molecule localization microscopy of cryosamples, and we describe the rationale behind its design. All specifications, including a detailed 3d model of the entire assembly, are freely available via ccb.lumc.nl/downloads-231.

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