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Cryo-FIB milling technology
Following localization by correlative microscopy, cryo-focused ion beam (cryo-FIB) microscopes can be used to prepare and isolate a region of interest within a sample. Since its inception, cryo-FIB technology has evolved significantly. Cryo-FIB systems based on liquid metal ion sources (LMISs) typically use a beam of gallium ions to scan across a frozen sample surface, removing surface atoms layer by layer in a process called sputtering or ion beam milling. This process can be used to create 100-200 nanometer thick electron-transparent in situ lamellae from vitrified cells by removing cellular material above and below a target site. The Thermo Scientific Aquilos 2 Cryo-FIB with the iFLM Correlative System is a dedicated solution for cryo-lamella preparation and targeting with fluorescence microscopy.
Plasma FIB technology
Plasma focused ion beam (PFIB) technology offers several types of ions for milling, including xenon, oxygen, argon, and nitrogen, as well as higher milling throughput compared to gallium FIB devices. The unique milling properties of the different ion species can produce high-quality lamellae without gallium implantation effects. The Thermo Scientific Arctis Cryo-PFIB and Thermo Scientific Hydra Bio Plasma-FIB support a range of applications in cryogenic conditions or at room temperature.
Lamellae preparation for cryo-electron tomography
Cryo-FIB technology has transformed life science research by providing access to biological samples preserved in a near-native state for structural analysis using cryo-electron tomography. In the cryo-tomography workflow, samples are rapidly frozen to turn water into amorphous (or vitreous) ice using a vitrification instrument, such as the Thermo Scientific Vitrobot Mark IV System, and then thinned to electron-transparency with an instrument like the Aquilos 2 Cryo-FIB, Arctis Cryo-PFIB, or Hydra Bio Plasma-FIB, before imaging on a cryo-transmission electron microscope (TEM) such as the Thermo Scientific Krios Cryo-TEM.
Visualizing 3D structures of biological specimens
Cryo-FIB technology can be used for three-dimensional visualization of biological samples preserved in a near-native state. A 3D volume is generated by sequential ablation of the sample with the ion beam and imaging of the freshly generated surfaces with the electron beam. The Hydra Bio Plasma-FIB supports a wide variety of life science applications to study the 3D structure of cells, tissues, and organisms. Users can choose to image cryogenically preserved or room-temperature specimens to ensure maximum flexibility for volume EM acquisitions.
Key features of cryo-FIB technology
Cryo-preparation station
Reliably load vitrified samples into the cryo-FIB and safely remove them after generating cryo-lamellae.
Cryo-liftout
Extract and transfer specific sample regions for further FIB-thinning and downstream imaging.
iFLM Correlative System
Use cryo-fluorescence microscopy directly within the cryo-FIB to localize structures of interest for cryo-lamella preparation.
Auto Slice and View Software
Collect 3D volumetric datasets of vitrified or resin embedded samples with automated serial milling.
Cryo-lamella preparation software
Automatically generate electron-transparent lamellae for high-resolution imaging using cryo-electron tomography.
For Research Use Only. Not for use in diagnostic procedures.