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The Linkam Cryo-Correlative Microscopy Stage

Jeff McGinn

As McCrone Microscopes & Accessories prepares to exhibit with Linkam Scientific at the 2024 Microscopy and Microanalysis conference in Cleveland, OH, we would like to highlight the Linkam cryo-correlative microscopy stage, CMS196V3.

Linkam's CMS196V3 cryo-correlative microscopy stage,
Linkam’s CMS196V3 cryo-correlative microscopy stage.

The motorized, encoded CMS196V3 is a cryo-correlative microscopy system that enables the full workflow of correlative light and electron microscopy (CLEM). While electron microscopy (EM) provides structural information at very high magnification and resolution, biological and chemical process information is limited due to staining and sample preparation techniques. Alternatively, fluorescence microscopy is very useful at detecting biological, chemical, and genetic events inside living cells. Cryo-CLEM allows for both low temperature fluorescence microscopy and cryo-EM by imaging the same sample location with both techniques and superimposing the combined information.

Click on any image to enlarge.

Correlative cryo-fluorescent light microscopy and cryo-electron tomography of WGA stained Streptomyces bacteria targeting cross-membrane lipid structures.
Correlative cryo-fluorescent light microscopy and cryo-electron tomography of WGA stained Streptomyces bacteria targeting cross-membrane lipid structures.
Cryo-fluorescence stage Linkam CMS196 and cryo-widefield fluorescence of mouse embryonic fibroblast cells on quantifoid gold finder grids, imaged at the Crick Institute, London.
Cryo-fluorescence stage Linkam CMS196 and cryo-widefield fluorescence of mouse embryonic fibroblast cells on quantifoid gold finder grids, imaged at the Crick Institute, London.
Overlay of super-res cryoCLEM and diffraction-limited cryoFLM images of U2OS cells, transfected with rsEGFP2-MAP2, over an 18.6 nm thick slice through the sample.
Overlay of super-res cryoCLEM and diffraction-limited cryoFLM images of U2OS cells, transfected with rsEGFP2-MAP2, over an 18.6 nm thick slice through the sample.

The CMS196V3 maintains the vitrified state of the sample by liquid nitrogen cooling, allowing for safe handling and transfer of your cryo samples. Up to three grids can be loaded into a specially-designed cassette, which is then transported from a plunge freezer in a small sealed container. The container is loaded into a pre-cooled dry sample loading chamber fitted on an upright fluorescent microscope. The cassette is then easily loaded onto the viewing bridge using special manipulation tools.

Highlights of the CMS196V3:

  • Self-contained cryo sample observation system which keeps the sample vitrified at a constant -195°C and avoids contamination
  • Automatic liquid nitrogen top off of the chamber
  • Integrated motorized XY stage with optical encoders for high precision movement and position readout of better than 1µm
  • Fully automated XY mapping
  • Mounting option for most research grade upright microscopes from all major manufacturers
  • Self-aligning magnetic sample cassette system for up to three sample grids
  • Support for EM-grids, Auto-Grids (FEI), Membrane carriers (Leica HPM100), Bessy grids, and planchettes
  • Sample cassette holder for safe and contamination-free sample loading, storage and transfer
  • Connects to plunge-freezing and high-pressure freezing sample preparation protocols
  • Integrated sensors for sample temperature monitoring
  • Full day of independent operation with optional 3L autofill LN2 Dewar
  • LCD display with button control, stand-alone joystick operation and full software control via USB interface
  • Automated heating “bake-out” cycle to drive out moisture

We are happy to run a demonstration for you – contact us today to learn more about this remarkable Linkam stage.

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