CMS196V⁴

Cryo-Correlative Microscopy Stage

CMS196V⁴ Cryo-CLEM stage overview

Linkam’s new CMS196V⁴ is a market-leading cryo-correlative microscopy system enabling cryo-fluorescence, the full workflow of Correlative Light and Electron Microscopy (CLEM) and Cryo-super resolution.

The CMS196V⁴ maintains the vitrified state of the sample by liquid nitrogen cooling and provides proven capabilities to safely handle and transfer your cryo-samples. These can then be imaged with optical microscopy whilst ensuring they remain free of contamination. The integrated, encoded and motorised XY stage enables the co-ordinate mapping required to find the same sample feature again on complementing modalities, for example TEM, FIB-SEM or X-ray systems.

Samples are typically mounted on EM grids or other carriers compatible with electron microscopy, allowing correlated light and electron microscopy. The CMS196V⁴ enables high resolution maps of the grid to be produced quickly and easily in combination with our NEXUS software.

Up to 3 grids can be loaded into a specially designed cassette which is transported from a plunge freezer in a small, sealed container. The container is loaded into a precooled dry sample loading chamber fitted on an upright fluoresence microscope. The cassette is then easily loaded onto the viewing bridge using special manipulation tools.

Originally developed in collaboration with Prof. Bram Koster's team at Leiden University Medical Centre (LUMC) in the Netherlands, the CMS196 series is the result of years of intensive research and development.

“The correlative stage was developed out of the necessity for a tool that uses fluorescence microscopy to help localise structures of interest in samples for high resolution cryo electron microscopy imaging. Over the years the system has evolved into an effective and efficient instrument that enables the imaging of cryo electron microscope images after light microscopy characterisation routinely.”

Prof. A.J. Koster - LUMC

CMS196V4 Key Features

Self-contained cryo sample observation system with automatic LN2 top-up of the sample chamber which ensures that the sample remains vitrified at liquid nitrogen temperatures (<-195°C) and does not become contaminated.
Short startup time of only 10 minutes with no mechanical connection.
Provides a highly stable platform allowing high precision automated mapping of the complete EM grid with 1 μm resolution.
Self-aligning magnetic sample cassette system ensures safe sample loading, transfer and storage. One cassette can hold up to 3 EM-grids. Dedicated Cassettes support standard EM grids, Autogrids/Clipped Grids, Planchettes, Bessy, membrane carriers(Leica HPF system),Si3N4 frames, CryoCapCell, JEOL CRYO ARM and custom sample holders.
Sample cassette holder for safe and contamination-free sample loading, storage and transfer.
Interchangeable cryo-bridge which allows multiple sample interface options and accommodates different custom needs.
Cordless, heated frost-free lid allows convenient viewing of the sample area.
Integrated LED condenser allows brightfield transmitted light, ideal for initial viewing of the grid and locating the sample.
Integrated sensors for real-time sample temperature monitoring.
The CMS196V⁴ can be configured and operated from the connected joystick touch panel. The joystick wheels allow intuitive sample navigation and swift sample review without a PC. The stage can also be controlled via USB and is supported by multiple software packages.
Support for EM grids, Autogrids (FEI, sometimes referred to as ‘clipped grids’), membrane carriers (Leica HPF system), Bessy grids, Planchettes, CryoCapCell and custom sample holders.
Sample cassette holder for safe and contamination-free sample loading, storage and transfer.
Motorised sample positioning can be operated from the connected joystick touch panel without a PC. The stage can also be controlled via USB for PC control and is supported by multiple software packages.
Connects to plunge-freezing and high-pressure freezing sample preparation protocols.
Built-in bake-out heating cycle to remove moisture in the sample chamber.
Optional objective lens’ heater enhances superior drift performance.
The CMS196V⁴ is compatible with most research-grade upright microscopes and high NA objectives. A quick-release mount further simplifies sample loading for some microscopes.

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Why Cryo-CLEM?

Electron microscopy (EM) provides structural information at very high resolution. However, it can give only restricted insight into biological and chemical processes due to limitations in staining and sample preparation processes. Fluorescence microscopy on the other hand is a very sensitive method to detect biological, chemical and genetic processes and events inside living cells.

Cryo-CLEM brings it all together: it is a technique that combines the individual advantages from both Fluorescence and EM by imaging the same sample location with both techniques, and superimposing the complementary information.

"We are using the cryo-correlative stage to screen cells trapped in a thin layer of ice prior to imaging the samples at synchrotrons in Oxford, Barcelona and Berlin. The team at Linkam Scientific have helped to drive forward our research, quickly adapting and refining the technology to deliver an elegant, flexible, user-friendly stage."
Dr. Lucy Collinson, Head of Electron Microscopy, The Francis Crick Institute, London, UK.

Imaging of vitrified samples under cryo-conditions

In order to make a biological sample compatible with the vacuum conditions found in EM and preserve the structural detail, samples are embedded in vitrified “glass-like” ice and need to be kept below -140°C. Any contact with moisture contained in the air has to be avoided since ice crystals would form immediately and contaminate the sample. Under cryo-conditions the fluorescence signals' structural detail is preserved and photobleaching is significantly reduced.

Read about the new Linkam CryoGenium plunge freezing system (under development)