The Linkam Cryo-Correlative Microscopy Stage

The Linkam CMS196V⁴ stage is a cryo-correlative microscopy system supporting Cryo-Correlative Light and Electron Microscopy (CLEM), a combination of fluorescence microscopy with high-resolution electron microscopy (EM). The CMS196V⁴ stage was designed specifically to solve the challenge of transferring vitrified EM grids from the fluorescent microscope into the cryo EM without devitrification and contamination through condensation.

While EM provides structural information at very high resolution, it can give restricted insight into biological and chemical processes due to limitations in staining and sample preparation processes. Alternatively, fluorescence microscopy is a very sensitive method to detect biological, chemical and genetic processes and events inside living cells. Cryo-CLEM brings together the dual advantages of low temperature fluorescence and cryo-EM by imaging the same sample location with both techniques, while also superimposing the complementing information. CLEM enables you to correlate the two different types of information on the exact same area of interest: cellular function from fluorescence microscopy and nanostructure from EM.

In CLEM, devitrification can be problematic since samples can transform from the desired amorphous state into a crystalline (ice) state, ultimately altering their native structures. The CMS196V⁴ maintains the vitrified state of the sample by liquid nitrogen cooling and provides proven capabilities to safely handle and transfer cryo-samples. These can then be imaged with optical microscopy while ensuring they remain free of contamination.

Samples are mounted on up to 3 EM grids and loaded into a variety of specially designed cassettes that are transported from a plunge freezer in a small, sealed container. The integrated and motorized XY stage provides mapping functionality to locate the same sample feature on light microscopes and other analytical techniques, such as TEM, FIB-SEM, or X-ray systems. Ultimately, high resolution maps are created in combination with Linkam NEXUS software.

Other unique features include:

• Autofill liquid nitrogen 33-liter dewar extends the unattended run time for up to 6 hours.

• Joystick touchscreen panel allows for intuitive sample navigation and review.

• Cordless heated magnetic sample chamber lid provides frost-free viewing of the sample area.

The Linkam CMS196V⁴ stage is being used by CLEM laboratories around the world, and more information can be found in these application notes.

Learn more about how the trusted advisors at McCrone Microscopes & Accessories can help with your CLEM research by requesting an online or in-person demonstration. Please contact us today.