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Cryo-plasma FIB sample preparation and imaging
Sample preparation is critical for all microscopy techniques and workflows. Thermo Scientific focused ion beam (FIB) and plasma focused ion beam (PFIB) microscopes can be used to prepare samples for a variety of imaging techniques, including cryo-electron tomography (cryo-ET), microcrystal electron diffraction (MicroED), and volume electron microscopy (vEM). These techniques allow us to image cellular ultrastructure in its native state at the nanometer scale and gain insights into the organization of cells within tissues at the micrometer to millimeter scales.
Sample preparation for cryo-electron tomography
Cryo-electron tomography (cryo-ET) is used to obtain 3D information from a variety of biological specimens, including purified molecules, protein complexes, viruses, bacteria, cells, and small organisms. Depending on the thickness of the specimen, a thinning step must be carried out using a cryo-FIB or PFIB microscope so that the sample can be penetrated by electron beams in the TEM.
Fluorescence localization with the iFLM Correlative System
Within the cryo-ET workflow, cryo-correlative microscopy can be used to identify structures of interest. A dedicated cryo-FIB stage keeps the sample in its vitrified state during fluorescence imaging. All Thermo Scientific cryo-FIBs and cryo-PFIBs can be equipped with an iFLM Correlative System, allowing samples to be imaged directly within the microscope without additional transfer steps to or from an external cryo-light microscope.
Left: Cryo-lamella prepared with Thermo Scientific Aquilos 2 Cryo-FIB. Right: Overlay of SEM image with fluorescence image obtained with iFLM Correlative System.
Sample preparation for microcrystal electron diffraction
MicroED is a technique used for structural determination of small molecules and proteins from nanometer-sized crystals. Small molecule crystals are usually dry and can be analyzed at room temperature. Protein crystals, however, are typically kept in water to retain their hydrated, native states. These samples are flash frozen (vitrified) to avoid sample damage due to crystalline ice formation. With MicroED, atomic details can be extracted from individual nanocrystals (<200 nm in size), even in a heterogeneous mixture. If the crystals are too thick, a cryo-FIB or cryo-PFIB microscope can be used to thin the crystal directly on the grid. This on-grid preparation method avoids further manipulation and transfer steps before imaging.
Sample preparation and imaging for volume electron microscopy
With modern 3D electron microscopy, the large volume of a sample can be captured by stacking sequential images so the true 3D structure of the specimen can be observed. FIB and PFIB microscopes are used to selectively remove material for sequential imaging with a SEM under cryogenic conditions or at room temperature.
Cryo-FIB and PFIB instruments
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Aquilos 2 Cryo-FIB |
Hydra Bio Plasma-FIB |
Arctis Cryo-Plasma-FIB |
| Dedicated cryo-FIB for cellular cryo-electron tomography sample preparation. | Versatile plasma-FIB for multi-application labs. | Automated cryo-plasma-FIB for throughput and connectivity. |
| The Thermo Scientific Aquilos 2 Cryo-FIB produces high-quality lamellae for cryo-ET. Key steps can be automated through user-friendly milling recipes. |
The Thermo Scientific Hydra Bio Plasma-FIB provides high-quality results for large volume microscopy and TEM lamella preparation, both at cryogenic and room temperatures. | The Thermo Scientific Arctis Cryo-Plasma-FIB automates high-throughput TEM lamellae production and features Autoloader connectivity for the cryo-ET workflow. |
Key features
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Key features
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Key features
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Applications
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Applications
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Applications
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| Datasheet: Aquilos 2 Cryo-FIB | Datasheet: Hydra Bio Plasma-FIB | Datasheet: Arctis Cryo-Plasma-FIB |
| See full product details | See full product details | See full product details |
For Research Use Only. Not for use in diagnostic procedures.