Sample Vitrification

Cells prepared by routine culture methods are surrounded by physiological solution and are grown on carbon-coated gold electron microscopy (EM) grids. Excess solution must be blotted away before the sample grid is plunge-frozen in a cryogenically cooled fluid, typically liquid ethane. The water in the sample freezes so rapidly that it does not crystallize, thus avoiding the molecular-scale disruption (by formed ice crystals) that would occur with a normal slow freezing process.

Vitrification with Vitrobot

Create high quality vitrified samples for single particle analysis or cryo-tomography research applications with Vitrobot. Vitrobot offers fully automated vitrification, fast and easy. It performs the cryo-fixation process at constant physical and mechanical conditions like temperature, relative humidity, blotting conditions and freezing velocity. This ensures high quality cryo-fixation results and a high sample preparation throughput prior to cryo-TEM observation.

The sample is transferred to a cryo-fluorescence light microscope, where structures of interest are identified by fluorescent tags. A dedicated cryo-LM stage keeps the sample in its vitrified state during cryo-fluorescence imaging.

Within the cryo-FIB, the LM data will be correlated using MAPS software, allowing quick identification of the region of interest with the cryo-FIB. On these regions of interest, a thin, uniform lamella must be prepared for cryo-ET. The dedicated cryo-FIB will allow for the sample to be milled at the proper temperature (approximately -170C), and it also prevents contamination of the sample.

Before milling, the sample is protected with a micrometer-thick metal layer (Pt) using the GIS. This layer prevents beam erosion on the sample. After milling, the lamella is sputtered with a nanometer-thin metal coating to render the sample conductive for TEM imaging. The cryo lamella is then transferred to a high-end TEM autoloader.

Aquilos Cryo-FIB

The Thermo Scientific™ Aquilos™ Cryo-FIB is the first cryo-DualBeam™ (focused ion beam/scanning electron microscope) system dedicated to preparation of frozen, thin lamella samples from biological specimens for high-resolution tomographic imaging in a cryo-transmission electron microscope (cryo-TEM).

The samples are placed within the autoloader in the TEM. Per cryo-lamella, a series of 2D images is acquired from different perspectives as the sample is tilted up to +/- 70 degrees.

Glacios Cryo-TEM

The new Thermo Scientific™ Glacios™ Cryo Transmission Electron Microscope (Cryo-TEM) delivers a complete and affordable Cryo-EM solution to a broad range of scientists. It features 200 kV XFEG optics, the industry-leading Autoloader (cryogenic sample manipulation robot) and the same innovative automation for ease of use as on the Krios G3i Cryo-TEM. The Glacios Cryo-TEM bundles all this into a small footprint that simplifies installation.

Krios G3i Cryo-TEM

The new Thermo Scientific™ Krios™ G3i Cryo Transmission Electron Microscope (Cryo-TEM) enables life science researchers to unravel life at the molecular level—easier, faster, and more reliably than ever before. Its highly stable 300 kV TEM platform and industry-leading Autoloader (cryogenic sample manipulation robot) are designed for automated applications, such as single particle analysis (SPA) and cryo-tomography. Designed-in connectivity ensures a robust and risk-free pathway throughout the entire workflow, from sample preparation and optimization to image acquisition and data processing.

Talos™ Arctica

The Talos Arctica is a 200kV FEG Transmission and Scanning Electron Microscope (STEM). It is a powerful, stable, and versatile system for delivering high-resolution 3D characterization of biological and biomaterials samples in cell biology, structural biology, and nanotechnology research. Talos enables scientists to quickly obtain better insight and understanding of macromolecular structures, cellular components, cells, and tissues in three dimensions.

In cases where multiple identical structures are present in the sample (e.g., pores in the cell’s nuclear membrane), the reconstruction and analysis software can combine data from individual structures, averaging out noise and increasing contrast to create a composite model with higher resolution. The operator can segment, color, and cross-section the model in a variety of ways to enhance its display and presentation.