Journal
/
Bioquímica
/
Preparation and Cryo-FIB micromachining of Saccharomyces cerevisiae for Cryo-Electron Tomography
JoVE Revista
Bioquímica
Author Produced
This content is Free Access.
JoVE Revista Bioquímica
Preparation and Cryo-FIB micromachining of Saccharomyces cerevisiae for Cryo-Electron Tomography

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

DOI:
10.3791/62351-v

•

09:06 min

•

November 20, 2021

•

, , ,
1Central European Institute of Technology,Masaryk University

Summary

Traducción Automática

Traducción Automática

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

Saccharomyces CerevisiaeCryo-electron TomographyCell CultivationSterile ConditionsLaminar Flow BoxYeast Extract MediumGlucoseExponential PhaseOD MeasurementGlycerolPlasma CleaningGlow DischargingVitrobotBlottingLiquid EthaneVitrification

Article

Embed

Añadir a la lista de reproducción

Usage Statistics

Videos relacionados

Quantitative Analysis of the Cellular Lipidome of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry

Utilization of Grafix for the Detection of Transient Interactors of Saccharomyces cerevisiae Spliceosome Subcomplexes

Labelling and Visualization of Mitochondrial Genome Expression Products in Baker’s Yeast Saccharomyces cerevisiae

Quantitative Metabolomics of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry

Fast Grid Preparation for Time-Resolved Cryo-Electron Microscopy

Preparing Lamellae from Vitreous Biological Samples Using a Dual-Beam Scanning Electron Microscope for Cryo-Electron Tomography

Assessment of Submitochondrial Protein Localization in Budding Yeast Saccharomyces cerevisiae

Micropatterning Transmission Electron Microscopy Grids to Direct Cell Positioning within Whole-Cell Cryo-Electron Tomography Workflows

Preparation of Nucleosome Core Particles Complexed with DNA Repair Factors for Cryo-Electron Microscopy Structural Determination

Author Spotlight: Advancing Chromatin Research and Overcoming Limitations with a High-Enrichment Locus-Specific Chromatin Isolation Protocol

Read Article