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Glacios 2 Cryo – Transmission Electron Microscopy (TEM)

A complete solution for high-resolution structure determination

The Thermo Scientific Glacios 2 Cryo-Transmission Electron Microscope (cryo-TEM) is a 200 kV microscope that allows you to easily collect near atomic data from a broad range of biological targets. Compared to the previous generation, the Glacios 2 Cryo-TEM delivers higher throughput and makes cryo-EM more accessible. It features an integrated Thermo Scientific Falcon 4i Direct Electron Detector, Thermo Scientific EPU Software, and a new full enclosure, which combine to enhance image quality, automate data acquisition, and simplify your work. The Glacios 2 Cryo-TEM is ideally suited for single-particle analysis, cryo-electron tomography (cryo-ET), and micro-electron diffraction (MicroED) applications.

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Cryo-TEM designed for high resolution and throughput

The Thermo Scientific Glacios 2 Cryo-TEM showcases many of the advances that have made high-quality analysis possible on 200 kV instruments. The Glacios 2 Cryo-TEM provides near atomic resolution. With improved hardware and a new enclosure that minimizes environmental influences, the Glacios 2 Cryo-TEM now has an information limit of 2.1 Å. Similarly, transmission loss due to ice growth has been reduced to 2% over 24 hours, helping you collect more high-resolution images with less effort.

Glacios 2 Cryo-TEM highlights

The combination of the Thermo Scientific Selectris X Energy Filter with the Thermo Scientific Falcon 4i Direct Electron Detector, the newly designed enclosure, and Thermo Scientific Smart EPU Software work together to guide the microscope and user to get the best possible results from their samples. The Glacios 2 Cryo-TEM in combination with the Thermo Scientific low-energy-spread cold FEG (E-CFEG), Thermo Scientific Selectris X Energy Filter, the Thermo Scientific Falcon 4i Direct Electron Detector, the newly designed enclosure, and Thermo Scientific Smart EPU Software work together to guide the microscope and user to get the best possible results from their samples with unprecedented resolution, speed, and ease of use at 200 keV.

The Thermo Scientific E-CFEG is a cold field emission gun with a narrow energy spread that results in higher contrast images and higher resolution. This add-on makes it possible to efficiently solve structures at resolutions of 2.5 Å or less.

Source
  • High-brightness field emission gun (X-FEG)
  • Low Energy Spread cold FEG (E-CFEG)
Accelerating voltage
  • Flexible accelerating voltage from 80–200 kV
Cryo-Autoloader
  • Cryo-Autoloader for automated and contamination-free loading of cassettes containing up to 12 Autogrids
Lenses
  • Automatic condenser, objective, and SA apertures
  • Symmetric constant power objective lens (minimize image aberrations and lens hysteresis during mode switching)
  • Symmetric constant power C-TWIN objective lens with wide-gap pole piece (11 mm)
Stage
  • Computerized 4-axis specimen stage with ±70° alpha tilt
  • Cryo-stage with single-axis holder
Imaging
  • Rotation-free imaging upon magnification changes
Installation requirement highlights
  • Environmental temperature: 18°C–23°C
  • Door height: 2.32 m (optionally reduced to 2.00 m)
  • Door width: 0.99 m
  • Ceiling height: 2.80 m
AFIS (aberration-free image shift)
  • Enhancing throughput with shorter relaxation times when moving coma-free between grid holes
FFI (fringe-free imaging)
  • Fringe-free imaging (FFI) for multiple image acquisitions per grid hole
Smart EPU Software
  • EPU 3 Software for automated sample screening and data acquisition (includes open application programming interface)
  • EPU Multigrid functionality (optional)
  • EPU Quality Monitor Software (optional)
  • Embedded CryoSPARC Live (optional)
Additional components
  • Primary control unit with two 24” monitors to be placed within 10 m of the column
Detectors (optional)
  • Falcon 4i Direct Electron Detector
  • Thermo Scientific Ceta-D Camera
  • Thermo Scientific Ceta-M Camera
  • HAADF STEM detectors*
  • On-axis BF/DF detectors*
  • *STEM is not compatible with FFI
Energy filter (optional)
  • Selectris and Selectris X Imaging Filters

High-resolution TEM results

Hardware optimization has been focused on improved specificity and sensitivity in electron detection. For instance, the Falcon 4i Direct Electron Detector enables high detective quantum efficiency (DQE) across the full spectral range, capturing small, flexible proteins as well as larger structures using fewer images. When coupled with Selectris or Selectris X Energy Filters, signal is limited to the zero-loss, elastically scattered electrons, producing high-contrast TEM data for higher throughput and higher resolution structures. The optional, integrated E-CFEG has a lower energy spread and better coherence than a traditional Schottky FEG, which enables higher contrast at high resolutions (≤2.0 Å) and to reach higher resolutions in less time.

High throughput cryo-TEM image acquisition

The integrated Falcon 4i Direct Electron Detector provides better image quality than its predecessors in less time. To further enhance productivity and image quality, the Glacios 2 Cryo-TEM offers fringe-free imaging (FFI) that eliminates fringes at the edge of the beam, a more stable stage, and faster automated data acquisition. Together, these improvements allow you to collect more images per foil hole and accelerate single particle analysis and cryo-ET. Adding the Selectris X Imaging Filter allows you to use a very stable narrow slit width (<10 eV) for an additional boost in contrast for single particle cryo-EM and cryo-ET, improving resolution and obtaining results faster with less data to maximize resolution.

Automated cryo-EM software maximizes ease of use and efficiency

Smart EPU Software enables faster and easier set up and processing, benefiting both expert and non-expert microscopists. Several critical set-up steps have been automated, such as alignment of the column and repair of system status, when possible. In addition, the software and various optional add-ons can help users get the most out of their microscope time and the best possible results.

Cryo-EM sample screening and workflow connectivity

Sample screening is an essential component of successful cryo-EM workflows, allowing verification of sample quality in terms of both biochemistry and vitrification. The Glacios 2 Cryo-TEM fits seamlessly into single particle analysis and cryo-ET workflows with convenient and contamination-free sample transfer throughout the workflow.

Smaller footprint

The hardware architecture of the Glacios 2 Cryo-TEM has been specifically designed with a smaller footprint and easier access path compared to other 200kV microscopes with automated sample loading systems.

High resolution single particle cryo-EM

With its high-resolution capabilities, the Glacios 2 Cryo-TEM is ideal for single particle analysis, enabling near-atomic structural determination of challenging proteins without the need for crystallization. Samples can be studied directly in solution. High-quality data collection is possible thanks to hardware improvements and the integration of innovative Thermo Scientific Smart EPU Software.

High throughput cryo-EM for drug discovery

Cryo-electron microscopy is rapidly drawing interest in structure-based drug discovery and design, since it can accurately and rapidly visualize to high resolutions the interactions between drug and receptor of a multitude of samples. The Glacios 2 Cryo-TEM can perform as a standalone tool for high resolution data collection, for example, CAK at 2.3 Å, but also serves as a powerful screening system that enhances subsequent 300 kV analysis. The Glacios 2 Cryo-TEM was used to determine over two dozen 3.5-4.5 Å kinase structures at ~1 structure per hour. This approach greatly improves throughput and productivity, optimizing the time spent on the higher-resolution Thermo Scientific Krios Cryo TEM and ensuring that the highest quality data is collected for only the most ideal samples. This methodology facilitates the application of cryo-EM to iterative structure-based drug design, which is becoming increasingly necessary to address a range of global health challenges.

Workflow using Glacios 2 Cryo TEM and Krios G4 Cryo TEM for evaluating CDK targeting compounds
Workflow using Glacios 2 Cryo TEM and Krios G4 Cryo TEM for evaluating CDK targeting compounds

Cryo-tomography

The Glacios 2 Cryo-TEM offers excellent cryo-electron tomography (cryo-ET) capabilities, providing nanometer-scale imaging of a cell’s interior in 3D and the visualization of protein complexes within their physiological environments. Using the Glacios 2 Cryo-TEM, tomography data can be obtained from whole bacterial cells or lamellae prepared with Thermo Scientific Aquilos, Arctis or Hydra Bio Cryo-focused ion beam (Cryo-FIB) instruments.

Microcrystal electron diffraction

The structural determination of small molecules and proteins can be challenging using standard methods such as X-ray crystallography. The Glacios 2 Cryo-TEM enables fast, high-resolution imaging using microcrystal electron diffraction (microED), where atomic details can be extracted from individual nanocrystals (<200 nm in size), even in a heterogeneous mixture. The Glacios 2 Cryo-TEM can be paired with the Thermo Scientific CETA-D Camera and the Thermo Scientific MicroED Package for a complete solution for MicroED.

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