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Introduction to cryo-EM
Sample preparation
Introduction to the microscope
Image formation and aberrations
Accelerator
Condenser lens and parallel illumination
Objective, lens, stage, and projection
Detectors and cameras
Single particle analysis data collection and EPU
Entire workflow uninterrupted
Getting started in cryo-EM
Detailed overview of the process to prepare samples for cryo electron microscopy analysis and the conditions that are needed to produce quality vitrified samples.
Review of different manufacturer grid boxes, the orientation of the grids, and how to remove the grids.
Demonstration of how to identify the different sides of grids- the copper and carbon sides of the grid.
Demonstration of how to properly hold grids with tweezers.
Demonstration and step by step instruction of how to glow discharge and plasma clean grids using a Pelco easiGlow.
Demonstration of the difference between a glow discharged and nonglow discharged grid.
Review of the tools needed for plunge-freezing samples with the Vitrobot System.
Walk through of the Vitrobot System connections and cables.
Demonstration of how to properly mount the Vitrobot System's humidifier.
Step by step demonstration of how to properly fill the Vitrobot System humidifier.
Walk through of the Vitrobot System humidifying chamber, including the different components and their purpose.
Demonstration of the operating software of the Vitrobot System.
Step by step instructions for replacing the Vitrobot System blot pads.
Demonstration on how to properly mount and unmount tweezers from the Vitrobot System.
Step by step demonstration of how to align the Vitrobot System tweezers between the blot pads before sample application.
Preparing the ethane gas source and safety considerations.
Demonstration of how to carefully fill the ethane cup with liquid ethane for plunge freezing with the Vitrobot System.
Step by step walk through of plunge freezing a sample with the Vitrobot System.
Demonstration of how to revive solid ethane to use it again for plunge freezing samples.
Walk through of the different sources of contamination in liquid nitrogen and recommended ethane quality.
Run through of the tools required to assemble standard plunge frozen EM grids into an autogrid cartridge.
Step by step instructions on how to properly insert C-clips into the insertion tool.
Run through of how to assemble autogrids step by step and the tools required without liquid nitrogen.
Step by step instructions on how to assemble autogrids with liquid nitrogen.
Walk through of the components needed to put the cartridges in the cassette.
Demonstration on how to properly load and mount a capsule onto the loading station and assembly of the cassette-loading station.
Walk through of the autoloader cassette and the different features of the cassette.
Demonstration on how to insert the autogrids into the cassette and optimal conditions for the cassette.
Run through of loading the autoloader cassette with autogrids without liquid nitrogen using the loading station.
Demonstration of loading autogrids into the autoloader cassette in the loading station with liquid nitrogen.
Walk through of the on-system display of the Talos Arctica Cryo-TEM for docking and undocking cassettes.
Demonstration of the capsule pin and its purpose for proper function of the autoloader.
Demonstration of docking a cassette into the autoloader on a Talos Arctica Cryo-TEM.
Demonstration of undocking a cassette out of the autoloader on a Talos Arctica Cryo-TEM.
Discussion of the biological hazards and precautions that can be used when working with hazardous agents.
Review of the hazards from compressed gases used in cryo electron microscopy and mitigation.
Discussion of the combustion hazards of ethane.
Demonstration of how flammable liquid ethane is.
Discussion and walk through of the hazards around large liquid nitrogen tanks.
Walk through of the hazards of different small liquid nitrogen Dewars.
Safety considerations when handling and working with liquid nitrogen.
Review of the safety hazards of handling ethane and the cold cup.
For Research Use Only. Not for use in diagnostic procedures.