Dual-Beam FIB/SEM: CrossBeam 350
The Zeiss CrossBeam 350 is a dual‑beam FIB/SEM instrument designed for multidisciplinary use in life sciences and materials science. It has cryogenic operation capability, facilitating the analysis of biological samples prepared using cryomethods (plunge freezing, high-pressure freezing, freeze substitution), ensuring excellent structural preservation.
The instrument enables integrated correlative observation with (cryo)confocal microscopy and includes micromanipulators and gas injectors for performing electrical measurements and preparing (cryo)lamellae that can subsequently be observed in Transmission Electron Microscopes (TEM), both at room temperature and under cryogenic conditions.
It also enables tomographic studies aimed at obtaining 3D structural and compositional information through EDS spectroscopy in a wide variety of samples. It is therefore an essential system in the field of Volume Electron Microscopy.
Instrument Features
The CrossBeam 350 is equipped with:
• An electron column based on Gemini® technology.
• A gallium ion column (Ion-Sculptor) operating across multiple voltage ranges.
• Two secondary electron detectors (Inlens and Everhart‑Thornley type chamber detector).
• Two backscattered electron detectors (annular retractable and Inlens).
• An UltimMax 100 EDXS detector (Oxford Instruments) with a minimum detection area of 100 mm².
• Two CCD cameras inside the chamber.
• Two gas injectors (tungsten and carbon).
• A Kleindiek Nanotechnik MM3A‑EM micromanipulator offering x, y, z and rotation movement, operable at room temperature and at liquid-nitrogen temperature.
• A cryo-preparation chamber with cryofracture capability (Quorum PP3010Z).
The system includes a single specimen holder that allows transfer from the confocal microscope to the FIB‑SEM, enabling the selection and observation of the same sample in a fully correlative manner, both at room temperature and at liquid-nitrogen temperature. This navigation is highly precise thanks to the navigation camera and the six-axis holder mechanics.
Technological Applications
The high versatility of this equipment enables new research avenues and technological applications in both materials science and life sciences. Examples include:
• Acquisition of high‑resolution images across a wide range of samples.
• Simultaneous observation with electron beam and ion beam.
• Preparation of high‑quality (cryo)lamellae in specific regions for subsequent TEM observation, including lift‑out, MEMS‑chip preparation for in situ studies, or lamella‑on‑grid preparation, at both room temperature and cryogenic temperature.
• Morphological and compositional characterization using the integrated detectors.
• Tomographic series acquisition with nanometric serial sectioning in biological and materials samples.
• Acquisition of 2D and 3D compositional information.
• Chemical analysis of microstructures.
• Creation or modification of structures using FIB and different gases.
• Image acquisition with spatial correlation to optical microscopy images.
• Full cryogenic workflow capability at liquid‑nitrogen temperature throughout the correlative process.
Funding
This equipment has been fully funded with European funds through the 2021 call for scientific and technical equipment acquisition of the Next Generation EU program, within the Recovery, Transformation and Resilience Plan, under project EQC2021‑007336‑P, with a grant of €1,228,000.
Location
The CrossBeam 350 dual-beam system is located within the Electron Microscopy and Related Techniques Unit, in the Advanced Microscopy building, and forms part of the singular infrastructures of the Barcelona node of the ICTS ELECMI.