ZEISS Crossbeam

Your FIB-SEM for High Throughput 3D Analysis and Sample Preparation

ZEISS Crossbeam

Your FIB-SEM for High Throughput 3D Analysis and Sample Preparation

Combine imaging and analytical performance of a high resolution field emission scanning electron microscope (FE-SEM) with the processing ability of a next-generation focused ion beam (FIB). You may be working in a multi-user facility, or an academic or industrial lab. Take advantage of ZEISS Crossbeam’s modular platform concept and upgrade your system with growing needs, e.g. with the LaserFIB for massive material ablation. During milling, imaging or when performing 3D analytics Crossbeam will speed up your FIB applications.

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Maximize Your SEM Insights
Increase Your FIB Sample Throughput
Experience Best 3D Resolution in Your FIB-SEM Analysis

The TEM lamella preparation workflow of ZEISS Crossbeam used by Benedikt Müller, University of Tuebingen, and Claus Burkhardt, NMI Reutlingen, to investigate the crystal structure of NanoSQUIDS.

Highlights

ZEISS Crossbeam with Gemini Optics.

Maximize Your SEM Insights

Take advantage of up to 30% better SEM resolution at low voltage using Tandem decel, a feature of the novel ZEISS Gemini electron optics.
Extract true sample information from your high resolution SEM images using Gemini electron optics.
Count on the SEM performance of your ZEISS Crossbeam for 2D surface sensitive images or when performing 3D tomography.
Benefit from high resolution, contrast and signal-to-noise ratios, even when using very low acceleration voltages.
Characterize your sample comprehensively with a range of detectors. Get pure materials contrast with the unique Inlens EsB detector.
Investigate non-conductive specimens undisturbed by charging artifacts.

Increase Your FIB Sample Throughput

Benefit from speed and precision of intelligent FIB scanning strategies for material removal and perform your experiments up to 40% faster than before.
The Ion-sculptor FIB column introduces a new way of FIB-processing: by minimizing sample damage you’ll maximize sample quality and perform experiments faster at the same time.
Manipulate your samples precisely and fast by using up to 100 nA current without compromising FIB resolution.
When preparing TEM samples use the low voltage capabilities of the Ion-sculptor FIB: get ultra-thin samples while keeping amorphization damage at a minimum.

The focused ion beam column, ZEISS Ion-sculptor, of ZEISS Crossbeam.

3D tomography of a solder, this image is part of a multi-modal workflow combining imaging and EDS analytics.

Experience Best 3D Resolution in Your FIB-SEM Analysis

Enjoy the benefits of integrated 3D analysis for EDS and EBSD investigations.
During milling, imaging or when performing 3D analytics Crossbeam will speed up your FIB applications.
Expand the capacity of your Crossbeam with ZEISS Atlas 5, the market-leading package for fast, precise tomography.
Perform EDS and EBSD analysis during tomography runs with the integrated 3D Analytics module of ZEISS Atlas 5.
Gain unique insights from best 3D resolution and leading isotropic voxel size in FIB-SEM tomography. Probe less than 3 nm in depth and produce surface sensitive, material contrast images using the Inlens EsB detector.
Save time by collecting your serial section images while milling. Achieve accuracy and consistency using trackable voxel sizes and automated routines for active control of image quality.

Crossbeam Family

Crossbeam 350

Leverage low vacuum operation and perform in situ experiments with outgassing or charging samples using the Variable Pressure mode. Achieve high quality imaging and high throughput enabled by the unique Gemini electron optics and the Ion-sculptor FIB.

Crossbeam 550

Perform your most demanding characterizations and choose the chamber size, standard or large, that best suits your samples. The Gemini 2 electron optics enables high resolution, even at low voltage and high current. It’s ideal for high resolution imaging at high beam current and for fast analytics.

Crossbeam laser

Your instrument for massive material ablation and preparation of large samples - the femtosecond laser on the airlock enhances in situ studies, avoids chamber contamination and is configurable as Crossbeam 350 and 550. Gain rapid access to deeply buried structures or prepare extremely demanding structures e.g. atom probe samples.

Correlative Cryo Workflow

This solution for TEM lamella preparation and volume imaging under cryogenic conditions enables imaging near-to-native state. Connect widefield, laser scanning, and focused ion beam scanning electron microscopy. Keep the flexibility of a multi-purpose FIB-SEM simultaneously.

Array of micropillars, produced with the LaserFIB, field of view 2.1 mm.

ZEISS Crossbeam Laser Workflow

How the LaserFIB Workflow Enhances Your High-resolution Imaging & Analysis

Rapidly access buried regions of interest, execute correlated workflows across multiple length scales, acquire better sample representativity with large-volume analysis, and perform 3D imaging and analytics. Add a femtosecond laser to your ZEISS Crossbeam and benefit from site-specific, ultra-fast sample preparation.

Watch this animation and discover how the LaserFIB workflow is used on an electronic sample. In this correlative experiment a defect was located non-destructively with XRM. Then the ROI was exposed with the femtosecond laser, fine polished by the FIB beam and finally analyzed with the SEM.

Gain rapid access to deeply buried structures
Prepare extremely large cross-sections up to millimeters in width and depth
Benefit from minimal damage and heat affected zones due to femtosecond laser pulses in a controlled vacuum environment
Perform laser work in a dedicated integrated chamber to maintain cleanliness of your FIB-SEM main chamber and detectors
Find your deeply buried regions of interest (ROIs) by correlation with previously acquired 3D X-ray microscopy datasets or other external data

1. Perform set-up steps for laser ablation

Load your sample onto the holder and transfer into the FIB-SEM main chamber
Import, overlay and align e.g. 3D X-ray data or 2D optical microscope images in ZEISS correlative workspaces
Find your ROI and acquire a reference image

2. Register SEM and laser coordinates

Scan the four sample holder fiducials with the SEM to lock sample and SEM coordinates
Transfer sample to integrated femtosecond (fs) laser chamber
Scan the four sample holder fiducials with the fs laser to lock sample and laser coordinates
SEM and laser coordinates are now aligned

Top down SEM view

Cross-section SEM view

3. Perform massive material ablation

Draw your laser pattern
Expose the laser pattern
Quickly remove massive material volumes with better than 2µm targeting accuracy

FIB-polished cross-section SEM view

Cross-section, detail showing the defect

4. Transfer the sample to the main FIB-SEM chamber to continue your FIB-SEM work

Details of the microstructures can already be observed
Perform FIB polishing as required for high-resolution imaging
Create TEM and atom probe samples with novel workflows
Rapidly optimize laser recipes with immediate SEM feedback

The Workflow for TEM Lamella Preparation

Just do it with high quality at high throughput

Navigate to your region of interest.

Navigate to your region of interest

TEM lamella preparation is essential for almost any FIB-SEM user. ZEISS offers an automated workflow for site-specific preparation. The resulting lamellae are ideally suited for high resolution TEM and STEM imaging and analysis at atomic resolution. Navigate to the specimen’s ROI, extract your TEM lamella including ROI from your bulk sample, perform the bulk milling or trenching step, and finalize the workflow with lift-out and thinning where appropriate.

1. Automated navigation to the specimen’s region of interest (ROI)

Begin the workflow without time-consuming search for the ROI
Use the navigation camera on the airlock to locate specimens
The integrated user interface makes it easy to navigate to your ROI
Benefit from the large, distortion-free field of view in the SEM

Lamella of a copper sample ready for lift out

2. Automated Sample Preparation (ASP) to prepare a lamella out of the bulk

Start the preparation with a simple three-step process: ASP
Define the recipe including drift correction, deposition and coarse and fine milling
The ion optics of the FIB column enables high throughput for the workflow
Duplicate the recipe and repeat as often as required in order to start a batch preparation

Part of the TEM lamella preparation workflow in a ZEISS Crossbeam

3. Lift out

Bring in the micromanipulator and attach the lamella to its tip
Cut out the lamella from the bulk
The lamella is then ready for lift out and can be transferred to a TEM grid

TEM lamella of a silicon sample after final thinning

4. Thinning: the final step is crucial, as it defines the quality of your TEM lamella

The instrument’s design allows you to reach a desired thickness of the lamella by enabling live monitoring of the thinning
Use two detector signals in parallel to judge lamella thickness and obtain reproducible end thickness on the one hand (with the SE detector) and to control surface quality on the other hand (with the Inlens SE detector)
Prepare high quality samples with negligible amorphization

TEM Lamella Preparation and Volume Imaging under Cryogenic Conditions

Components of the ZEISS Cryo Accessory Kit

Cryogenic microscopy allows the examination of cellular structures in their near-to-native state. However, users face complex challenges, such as preparation, devitrification, ice contamination, loss of samples or correlation across imaging modalities. ZEISS Correlative Cryo Workflow connects widefield, laser scanning, and focused ion beam scanning electron microscopy in a seamless and easy-to-use procedure. Hardware and software are optimized for the needs of correlative cryogenic workflows, from localization of fluorescent macromolecules to high-contrast volume imaging and on-grid lamella thinning for cryo electron tomography.

Imaging the near-to-native state

Seamless cryogenic workflow across multiple modalities
Sample protection against devitrification and ice contamination
High-resolution fluorescence imaging
High-contrast volume imaging and 3D reconstruction
Targeted on-grid lamella thinning for cryo TEM applications
Multipurpose use for cryogenic and room temperature applications

A simplified workflow to help you focus on your research

With ZEISS Correlative Cryo Workflow, you master the challenging combination of different imaging modalities under cryo conditions. The workflow solution connects light and electron microscopy, enabling volume imaging and efficient production of TEM lamellae. Dedicated accessories simplify the workflow and facilitate a safe transfer of cryo samples between the microscopes. Data management is assured by ZEN Connect, which keeps your data in context throughout the workflow. A series of processing tools help you enhance the imaging results.

Double-labelled yeast cells (CNM67-tdTomato and NUP-GFP).

Superior components to give you best-in-class data quality

Thanks to cryo-compatible objectives and the high sensitivity of the Airyscan detector, ZEISS LSM systems enable you to detect proteins and cellular structures at high resolution while gentle illumination and constant low temperatures prevent your samples from devitrification. The ZEISS Crossbeam FIB-SEM lets you enjoy high-contrast volumetric imaging – even without heavy metal staining applied to your samples. Both modalities provide valuable functional and structural information that can give you a thorough understanding of ultrastructure, whether or not you follow up with TEM studies.

Core Imaging Facility with Cryo equipment

Multipurpose solutions to maintain your imaging facility’s productivity

Unlike other solutions, the ZEISS microscopes involved in the workflow can be used not only for cryogenic microscopy, but also for room temperature applications, which is particularly advantageous when the microscopes are not being fully utilized for cryogenic experiments. Converting the instruments from cryogenic to room temperature usage is done quickly and doesn’t require technical expertise. This flexibility gives users more time for their experiments. Imaging facilities benefit from better utilization and a faster return on investment.

The Technology Behind ZEISS Crossbeam

SEM Electron Optics

Choose between Two Columns

The FE-SEM column of ZEISS Crossbeams is based on Gemini electron optics as all ZEISS FE-SEMs. Decide on the Gemini VP column of Crossbeam 350 or the Gemini II column of Crossbeam 550.

Field emission SEMs are designed for high resolution imaging. Key to the performance of a field emission SEM is its electron optical column. Gemini technology comes with all ZEISS FE-SEMs and FIB-SEMs: it is tailored for excellent resolution on any sample, especially at low accelerating voltages, for complete and efficient detection, and ease-of-use.

Gemini Optics is Characterized by Three Main Components

The Gemini objective lens design combines electrostatic and magnetic fields to maximize optical performance while reducing field influences at the sample to a minimum. This enables excellent imaging, even on challenging samples such as magnetic materials.
Gemini beam booster technology, an integrated beam deceleration, guarantees small probe sizes and high signal-to-noise ratios.
The Gemini Inlens detection concept ensures efficient signal detection by detecting secondary (SE) and backscattered (BSE) electrons in parallel minimizing time-to-image.

Benefits for Your FIB-SEM Applications

Long-term stability of the SEM alignment and the effortless way it adjusts all system parameters such as probe current and acceleration voltage
Achieve distortion-free, high resolution imaging even over large fields of view with the help of the near magnetic-field free optics
Tilt the specimen without influencing the electron optical performance

ZEISS Crossbeam with Gemini I SEM Column

ZEISS Crossbeam 350: Gemini column with single condenser, two Inlens detectors and VP capability.

ZEISS Crossbeam with Gemini II SEM Column

ZEISS Crossbeam 550: Gemini II column with double condenser and two Inlens detectors.

Crossbeam 350 with Gemini I VP

Maximum sample flexibility in multi-purpose environments
In situ experiments with outgassing or charging samples
Unique Gemini material contrast with the Inlens EsB detector

Crossbeam 550 with Gemini II

High resolution even at low voltage and high current thanks to the double condenser system
More information in less time with high resolution imaging and fast analytics
Unique topographical and material contrast with simultaneous Inlens SE and EsB imaging

Gemini Novel Optics

Profit from Surface Sensitive Imaging

Today’s SEM applications demand high resolution imaging at low landing energy as a standard. It is essential for:

beam sensitive samples
non-conductive materials
gaining true sample surface information without undesirable background signal from deeper sample layers

The novel Gemini optics are optimized for resolutions at low and very low voltages and for contrast enhancement.
Technological characteristics are the high gun resolution mode and the optional Tandem decel.

The high gun resolution mode results in minimized chromatic aberration thanks to of a reduction of primary energy width by 30%.

ZEISS Crossbeam 550 Objective with Tandem decel

Tandem decel optional sample biasing up to 5 kV further improves the excellent imaging capabilities at low voltages.

Tandem decel, now introduced to ZEISS Crossbeam 350/550, can be used in two different modes:

Tandem decel, a two-step deceleration mode, combines the beam booster technology with a high negative bias voltage that is applied to the sample: the electrons of the primary electron beam are decelerated, thus the landing energy is effectively reduced
Apply a variable negative bias voltage between 50 V and 100 V. One application mode enhances the contrast of your images
Apply a negative bias voltage between 1 kV and 5 kV and improve the low kV resolution of your images

FIB-SEM Technology

Discover a new way of FIB processing

The Ion-sculptor FIB column speeds up your FIB work without compromising machining precision and lets you benefit of its low voltage performance for any sample.

ZEISS Crossbeam 550 with a Gemini II column incl. double condenser and two Inlens detectors and a FIB-column arranged at an inclination angle of 54°.

The ZEISS Crossbeam Family carries the next-generation focused ion beam column, Ion-sculptor, featuring high currents for high throughput and excellent low voltage performance for high sample quality.

Maximize sample quality by using the low voltage capabilities of the Ion-sculptor FIB column
Minimize amorphization of your specimens and get the best results after thinning
Get precise and reproducible results with maximum stability
Accelerate your FIB applications with fast probe current exchanges
Perform high throughput experiments thanks to beam currents of up to 100 nA
Achieve exceptional FIB resolution of less than 3 nm
The Crossbeam family comes with automatic FIB emission recovery for long-term experiment

Applications

Materials Science

Nanomaterials
Live imaging of FIB-milling a spiral in silicon. Imaged with the SEM using an Inlens detector.

A)

Close-up of cross-section shows surface information on an Inlens SE image.

B)

The distribution of lanthanum (red) and manganese (green) is derived from an EDS map

Energy Materials
Lithium ion battery example, product key featuring cross-sectioning & 3D Tomography and 3D Analysis LiMn2O4 cathode material of a lithium ion battery. Close-up of cross-section shows surface information on an Inlens SE image ^A). The distribution of lanthanum (red) and manganese (green) is derived from an EDS map ^B).

Engineering Materials
H-bar lamella preparation by fs-laser on a copper semi-circle grid. The left lamella is 400 μm wide, 215 μm deep and has a thickness of about 20 μm at the top. It was cut by the laser in 34 s. The amount of material that needs to be removed by FIB for final thinning is significantly reduced.

Array of compression testing pillars in high entropy alloy, machined fully automatically

Engineering Materials
Key feature fs laser machining: Array of compression testing pillars in high entropy alloy, machined fully automatically.

Electronics & Semiconductor

3D-NAND – FIB-SEM Tomography
FIB-SEM tomography dataset acquired from a commercially purchased 3D NAND sample. Sample was depackaged and mechanically polished down to the topmost word line. Data acquisition was done on ZEISS Crossbeam 550 using ZEISS Atlas 3D. Voxel size 4 x 4 x 4 nm³.
Left: 3D rendering of the complete 2 x 4 x 1.5 µm³ volume.
Middle: Virtual sub-volume of 2 x 1.5 x 0.7 µm³ size, extracted from the dataset at the transition region of upper to lower deck.
Right: Single horizontal slice taken from the volume, showing a top-down view of a word line.

Power Electronics – Insulated Gate Bipolar Transistor
Insulated Gate Bipolar Transistor (IGBT) device analysis. The analysis was performed entirely on ZEISS Crossbeam 550.
First, a FIB cross-section across the gate electrode was cut, exposing irregularly distributed dark features (top).
Second, a lamella was prepared from the left-hand part of that cross section, and imaged by 30 kV STEM-in-SEM. The brightfield STEM image shown here reveals that the features are crystalline precipitates (bottom).
Third, EDS elemental mapping of the lamella showed that the precipitates are silicon (right).

Crossbeam laser FIB-SEM provides fast, high-quality cross sections of Cu-pillar microbumps buried 760 µm deep with total time to results of <1 hour.

3D Stacked Die Interconnect
Crossbeam laser provides fast, high-quality cross sections of Cu-pillar microbumps buried 760 µm deep with total time to results of <1 hour.
Left: 3D integrated circuit (IC) flip chip prepared for microbump imaging with laser ablation and FIB polishing.
Right: 25 µm diameter microbump image acquired with backscattered electrons.

Life Sciences

Cell Biology – HeLa Cells
Investigation of different cell compartments in single cells.
Individual HeLa cells were grown in culture dishes, chemically fixed
and resin-embedded in EPON. Voxel size 5 × 5 × 8 nm,
Inlens EsB detection, 1400 sections. 3D visualization with
Dragonfly Pro, ORS. Courtesy: A. Steyer and Y. Schwab, EMBL,
Heidelberg, DE.

Developmental Biology – C. elegans
Understanding the morphology of a whole organism in 3D with the highest resolution and reliability. The data set shows a arge 3D volume of C.elegans consisting of 10.080 z-sections at 5 x 5 x 8 nm pixel size. The nematode was high pressure frozen and freeze-substituted in EPON. Even the smallest structures inside the worm can be identified very easily.
Courtesy: A. Steyer and Y. Schwab, EMBL Heidelberg, DE; and S. Markert and C. Stigloher, University of Wuerzburg, DE.

Neuroscience – Brain Sections
Large area milling and imaging of a brain section with the
3D module of ZEISS Atlas 5. High current allows fast milling and
imaging of large fields of view up to 150 μm in width. The depicted
brain image has a field of view of 75 μm in width and
was milled with a beam current of 20 nA. Courtesy: C. Genoud,
FMI Basel, CH.

Microbiology - Trypanosoma
Ultrastructural investigation of the parasite Trypanosoma brucei.
The cells are high pressure frozen and freeze-substituted in EPON. Acquisition of 800 z-sections which corresponds to ~ 8 μm thickness in z; pixel size in x/y is 5 nm. Sample courtesy: S. Vaughan, Oxford Brookes University, Research Group ’Cell biology of Trypanosomes’, UK.

Accessories

Lithium-ion Battery

Visualization and Analysis Software

ZEISS recommends Dragonfly Pro from Object Research Systems (ORS)
An advanced analysis and visualization software solution for your 3D data acquired by a variety of technologies including X-ray, FIB-SEM, SEM and helium ion microscopy.

Available exclusively through ZEISS, ORS Dragonfly Pro offers an intuitive, complete, and customizable toolkit for visualization and analysis of large 3D grayscale data. Dragonfly Pro allows for navigation, annotation, creation of media files, including video production, of your 3D data. Perform image processing, segmentation, and object analysis to quantify your results.

Introducing ToF-SIMS enables High Throughput in 3D Analysis

Add the ToF-SIMS (time of flight secondary ion mass spectrometry) spectrometer to your Crossbeam 350 or Crossbeam 550 and analyze trace elements, light elements (e.g. lithium), and isotopes. Profit from sensitive and comprehensive analyses in 3D. Perform elemental mapping and depth profiling. Benefit from parallel detection of atomic and molecular ions down to the ppm level, achieve resolutions better than 35 nm in lateral direction and 20 nm in depth. Retrieve any signal from the ROI post-mortem.