Hydra Bio Plasma-FIB
Volume EM for cryo and resin-embedded samples
The Thermo Scientific Hydra Bio Plasma-FIB enables comprehensive exploration of biological samples, from tissue structures to proteins, through volume electron microscopy at both cryogenic and room temperatures, as well as cellular cryo-tomography. Optimal imaging quality starts with precise sample preparation. The Hydra Bio Plasma-FIB supports this by offering a choice of four ion species—argon, nitrogen, oxygen, and xenon—that are compatible with all commonly used embedding media and preparation techniques, such as high-pressure tissue freezing and critical point drying.
The Hydra Bio Plasma-FIB is designed to support a variety of workflows, including high-resolution serial PFIB imaging, large-area serial PFIB imaging, array tomography, correlative light and electron microscopy (CLEM), and high-throughput lamella preparation for cryo-tomography.
Features
Volume electron microscopy at cryogenic temperatures
Volume imaging for new studies for frozen-hydrated specimens with high-resolution serial PFIB milling and imaging in native state with excellent contrast.
Volume electron microscopy at room temperature
Multiple ion species (Xe, Ar, O, and N) for a range of sample materials and the Thermo Scientific Spin Mill Bio Method, an exclusive technique for large-area planar milling that yields similar sized imaging areas as microtome slicing.
Cryo-lamellae for cryo-electron tomography
Prepare lamellae from within large (HPF) volumes with plasma FIB, CLEM localization through integrated light and electron imaging with Thermo Scientific iFLM Correlative System and cryo-lift-out with Thermo Scientific Easylift Cryo Lift-Out System.
3D imaging at cryogenic temperatures
The Hydra Bio PFIB Cryo Package includes a fully integrated 360° rotation cryo-stage to maintain sample temperature below –170°C and a cryo-sample loading system to ensure that samples remain at cryo temperatures during transfer in and out of the instrument. With the excellent contrast of the Hydra Bio SEM column, uncover sub-cellular details on high pressure frozen and plunge frozen samples. No staining is required. Cryo-fixation avoids the artifacts that are usually associated with classic sample preparation steps with excellent contrast and minimal charging artifacts. Integrated Micro-Sputter Coater reduces charging under cryogenic conditions. Dedicated cryo-lamella preparation software supports your workflow with Thermo Scientific AutoTEM Cryo Software or cryo-volume imaging with Thermo Scientific Auto Slice & View Software.
Multiple ion options for optimized PFIB results
The Hydra Bio Plasma-FIB features a unique multiple ion species PFIB column with four ion species (Xe, Ar, O, N) with a patented, automated, and fast switching capability (<10 minutes). For example, O+ ion-plasma provides superior data acquisition efficiency and image quality for samples embedded in either epoxy- or acrylic-based resins. Unlike Ga-based FIBs, curtain-free surfaces are easily generated for a wide range of materials such as LR-White, HM20, EPON and Quetol resins.
Spin Mill Bio Method for millimeter-scale planar milling and imaging
The unique Thermo Scientific Spin Mill Bio Method on the Hydra Bio Plasma-FIB provides large-area planar milling up to 1 mm in diameter, with a geometry like microtome-based serial block face imaging but at a slice thickness as small as 5 nm. Uncover and visualize large sample areas with Spin Mill Bio Method, then focus in on your regions of interest. It is also ideal to prepare clean and smooth surfaces. The Spin Mill Bio Method is fully automated and easy to set up using guided software. Multiple areas for image acquisition within one experiment can be selected. Each area of interest can be imaged at different imaging settings based on the specificity of the experiment. Sparse features can easily be identified, and statistically relevant 3D data can be collected from multiple areas.
Integrated light microscope for correlative imaging
The integrated iFLM Correlative System combines fluorescence and electron imaging with ion milling and guided lamella production and supports correlative (CLEM) workflows. Identify, target, and confirm regions of interest without the hassle of transferring between instruments. A suite of shuttles is available, including a Leica External CLEM shuttle, to support grid or common HPF specimen formats. The iFLM Correlative System has been optimized for low distortions across a large field of view and exceptionally low mismatch between color channels. For external data sources Thermo Scientific Maps Software is available for correlation.
Specifications
• High performance PFIB column with Inductively Coupled Xe+ Plasma (ICP)
Ion beam current range: 1.5 pA to 2.5 µA
Accelerating voltage range: 500 V – 30 kV
Maximum horizontal field width: 0.9mm at beam coincidence point
• Ion beam resolution at coincident point
<20 nm at 30 kV using preferred statistical method
<10 nm at 30 kV using selective edge method
• Elstar extreme high-resolution field emission SEM column with:
- Immersion magnetic objective lens
- High-stability Schottky field emission gun to provide stable high-resolution analytical currents
- UC+ monochromator technology
• At optimum WD
• 0.6 nm at 15 kV
• 1 nm at 500 V
• At coincident point
• 0.7 nm at 15 kV
• 1.2 nm at 1 kV
• Elstar in-lens SE/BSE detector (TLD-SE, TLD-BSE)
• Elstar in-column SE/BSE detector (ICD)*
• Everhart-Thornley SE detector (ETD)
• IR camera for viewing sample/column
• High-performance ion conversion and electron (ICE) detector for secondary ions (SI) and electrons (SE)
• In-chamber Nav-Cam sample navigation camera*
• Retractable low voltage, high contrast directional solid-state backscatter electron detector (DBS)*
• Integrated beam current measurement
High-precision, 5-axis motorized stage with XYR axis, piezo-driven
- XY range: 150 mm
- Z range: 10 mm
- Rotation: 360° (endless)
- Tilt range: -38° to +60°
- XY repeatability: 1 μm
- Max sample height: Clearance 55 mm to eucentric point
- Max sample weight at 0° tilt: 500 g (including sample holder)
- Max sample size: 150 mm with full rotation (larger samples possible with limited rotation)
- Compucentric rotation and tilt