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CLEM — Cryo-fluorescence + cryoTEM

Correlative Light Electron Microscopy (CLEM) at AMF

A correlative cryo-imaging workflow combining a Zeiss AxioImager upright fluorescence microscope with a Linkam cryo-stage for cryo-fluorescence, and a Thermo Fisher Glacios cryo-transmission electron microscope (cryoTEM) for high-resolution structural imaging on the same vitrified sample. The Linkam stage accepts plunge-frozen TEM grids and autoloader cartridges directly, so grids can move between instruments under liquid nitrogen using a standard cryo-grid container, without remounting. Hosted in collaboration with the group of Lorenzo Albertazzi and the Center for Multiscale Electron Microscopy (CMEM), led by Heiner Friedrich.

Configuration — cryo-fluorescence side

  • Stand: Zeiss AxioImager upright fluorescence microscope
  • Cryo stage: Linkam cryo-correlative stage, accepts TEM grids and Glacios autoloader cartridges, LN2 cooled
  • Illumination: Zeiss Colibri LED for routine epi; custom high-power epi-illumination module via Coherent OBIS laser on the left side port (0.5 mW – 140 mW range) — built in-house by Mrigank Verma (AMF Imaging Scientist)
  • Structured illumination: Apotome (off during high-power laser epi)
  • Objectives: 10×, 50× (cryo-compatible with stage lid sleeve), 100×
  • Software: Zen (Zeiss); Coherent desktop software for the OBIS laser

Configuration — cryoTEM side

  • Instrument: Thermo Fisher Glacios cryoTEM, hosted at CMEM (Helix)
  • Sample format: Plunge-frozen EM grids in autoloader cartridges, transferable from the Linkam stage to Glacios under LN2
  • Workflow: Locate features by cryo-fluorescence on the AxioImager, transfer the cartridge under LN2 to Glacios for cryoTEM imaging at the same coordinates

Located at

Cryo-fluorescence (Zeiss AxioImager + Linkam) at AMF, ICMS. Glacios cryoTEM at CMEM, Helix building. Sample transfer between buildings under LN2 in a standard cryo-grid dewar.

Status

Operational. The custom Coherent OBIS epi-illumination module was completed in 2025 and is in routine use; technical questions about the custom light path go to Mrigank Verma. Day-to-day cryo workflows are coordinated with Mallika Chaukar (Albertazzi group). Vendor support: ZEISS Research Microscopy Solutions (Sanam Foroutanparsa, Edwin Lamers).

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Questions or training? Ask the super-user

Mallika ChaukarSuper-user · CLEM (cryo-fluorescence)
✉ m.chaukar@tue.nl

For general facility access or new project scoping, contact superresolution@tue.nl.