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Biochemistry
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Preparation and Cryo-FIB micromachining of Saccharomyces cerevisiae for Cryo-Electron Tomography
 

Preparation and Cryo-FIB micromachining of Saccharomyces cerevisiae for Cryo-Electron Tomography

Article DOI: 10.3791/62351-v 09:06 min November 20th, 2021
November 20th, 2021

Summary

We present a protocol for lamella preparation of plunge frozen biological specimens by cryo-focused ion beam micromachining for high-resolution structural studies of macromolecules in situ with cryo-electron tomography. The presented protocol provides guidelines for the preparation of high-quality lamellae with high reproducibility for structural characterization of macromolecules inside the Saccharomyces cerevisiae.

Tags

Preparation Cryo-FIB Micromachining Saccharomyces Cerevisiae Cryo-Electron Tomography Yeast Cultivation Sterile Conditions Laminar Flow Box Glucose Yeast Colony Growth Medium Cell Culture Incubator Exponential Phase Physical Density Holding Meter Glycerol Blanche Freezing Electro-microscopic Grids Plasma Cleaning
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