Aquilos Cryo-FIB for Life Sciences

Prepare cryo-lamellas for cryo-electron tomography

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). Cryo-electron tomography’s ability to visualize structures in their native context allows researchers to observe functional relationships and interactions with other components in the cellular environment. This technique promises to become an important tool for scientists seeking a better understanding of living systems at the molecular level.

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Key benefits

Produce cryo-lamellas for cryo-tomography using autoloader TEM systems.Cryo-dedicated hardware inside the DualBeam system chamber ensures optimal sample preparation while maintaining cryo conditions. Cryo specimens are mounted in dedicated Cryo-FIB AutoGrids which can be loaded into AutoLoader TEM systems.

Create thinner cryo-lamella without cutting artifacts. Ion beam milling enables creation of compression-free cryo-lamella samples for TEM tomography imaging. This method avoids artifacts such as mechanical compression inevitable for mechanical cryo-sectioning approaches using cryo-ultramicrotomes.

Improve sample preparation precision. Guided user software and Thermo Scientific™ MAPS™ correlation software makes the Aquilos Cryo-FIB easy to use, even for inexperienced users.

Assembly of a Nucleus-Like Structure During Viral Replication in Bacteria

Courtesy of Vorrapon Chaikeeratisak, Katrina Nguyen, Kanika Khanna, Axel F. Brilot, Marcella L. Erb, Joanna K. C. Coker, Anastasia Vavilina, Gerald L. Newton, Robert Buschauer, Kit Pogliano, Elizabeth Villa, David A. Agard, Joe Pogliano

In press. Science, 2017. DOI: 10.1126/science.aal2130

Enhanced sample integrity

The ability to properly prepare thin lamellas from vitrified samples, while at the same time, maintaining the necessary temperature, vacuum and transfer conditions, has been a significant challenge for researchers. To remove that impediment and make tomography accessible to a much broader community of researchers and scientists, the Aquilos Cryo-FIB features an integrated, fully rotatable cryo-stage and adjacent cryo-hardware that protects the frozen-hydrated samples from contamination, ensuring that delicate life science cryo-samples will be kept at vitreous temperatures ( < -165°C) at all times.

Integrated into cryo-tomography workflow

The Aquilos Cryo-FIB is specifically designed to complete our cryo-tomography workflow:

This workflow starts with vitrifying (ultra-fast freezing) the sample in an automated device (Thermo Scientific™ Vitrobot™).
An optional screening step follows, using cryo-fluorescence microscopy to identify and locate fluorescently-labelled features of interest. Any type of cryo-fluorescence data can be correlated using Thermo Scientific™ MAPS™ software.
The frozen sample is then moved to the Aquilos Cryo-FIB, which creates the thin cryo-lamella within a frozen-hydrated cellular sample.
The thin lamella (<200 nm) is then transferred to a cryo-transmission electron microscope (cryo-TEM) system such as the Thermo Scientific™ Krios™ or Thermo Scientific™ Talos Arctica™ Cryo-TEM for high-resolution imaging—all within our cryo-sample-transfer hardware, which protects and preserves the frozen sample.
The cryo-TEM acquires multiple images from different perspectives, from which a computer creates a 3D tomogram. Advanced image processing using subtomogram averaging allows high-resolution information to be extracted from the tomogram, modeling cellular proteins and features.

Featured Document

Aquilos Cryo-FIB Datasheet

The Thermo Scientific Aquilos Cryo-FIB is a cryo-dedicated DualBeam system that delivers optimal sample preparation for high-end cryo-TEM tomography. The Aquilos completes our cryo-electron tomography workflow, allowing researchers to reliably create samples with precisely controlled thickness with minimal investment in time and effort.

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Watch how, after vitrification of the sample, a DualBeam FIB-SEM is used for in situ lamella milling. This on-the-grid thinning prepares the sample for automated loading into a cryo-TEM for imaging.

Visualization of the nuclear periphery of a HeLa cell revealed by cryo‐electron tomography. The interphase HeLa cellwas thinned by cryo‐focused ion beam (cryo‐FIB) milling. The 3D segmentation of the tomographic data shows microtubules (orange), actin filaments (yellow), endoplasmic reticulum (green), nuclear pore complexes (light blue),large (dark blue) and small (brown) ribosomal subunits.

Data courtesy of Dr. J. Mahamid, Department of Molecular Structural Biology, Max Planck Institute for Biochemistry,Martinsried, Germany.

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