LUMC Light and Electron Microscopy Facility

Equipment

Light Microscopy

Confocal Laser Scanning Microscopes

We host a broad range of high-end confocal laser scanning microscopes. For live cell imaging we have two Leica SP8 with a white light laser (WLL) source, this allows you to perfectly match the spectral properties of any fluorophore excitable in the visible spectral range. Furthermore, we have a Leica SP5 STED for super-resolution microscopy capable of resolving sub-cellular details below 80 nm.

For intravital imaging we host a Zeiss LSM 7 MP at the Centre for Pre-clinical Imaging.
 

Currently we have the following confocal setups:

Inverted:

  • Leica SP8 WLL
  • Leica SP8 WLL
  • Leica SP5 STED
  • Leica SP5

Upright:

  • Leica SP8
  • Zeiss LSM 7 MP

Widefield Microscopes

Whereas confocal microscopes excel in 3D imaging, widefield microscopy is unrivalled in terms of speed and sensitivity. We have two (inverted) microscopes dedicated for live cell imaging, three upright microscopes for (up to 8 channels) multichannel fluorescence, and one microscope equipped with a color camera for brightfield imaging.

Inverted:

  • Leica AF6000
  • Leica DM6000

Upright:

  • Leica DM6B
  • Leica DM5500
  • Leica DMRA
  • Leica DMRB (color camera)

Cryo Fluorescence Microscopes

For cryo correlative microscopy we have two fluorescence microscopes equipped with special cryo-stages. One system is specifically set up for automated cryo-CLEM imaging, capable of performing automated multi-color, z-stack recording and recording tile-scan overviews. The other system is specifically used to develop super-resolution light microscopy on cryo-samples.

  • Zeiss Axio Image M2 with Linkam CMS196M cryo-stage
  • Leica DM RXA with Linkam THMS 600 freezing stage

Stereo Microscopes

  • Leica M165 FC

Slidescanners

  • 3DHistech Pannoramic 250

Electron Microscopy

Our facility houses a full range of electron microscopy infrastructure: tools for sample preparation, several electron and light microscopes, and hardware and software for computing and visualization.

Microscopy

The electron microscopy department has several electron microscopes and has access to two Titan Krios microscopes at the NeCEN facility. 

  • Zeiss Gemini SEM 300

The Zeiss Gemini SEM is a Field Emission Scanning Electron Microscope for imaging of micro structures. The microscope is equipped with the Gatan 3View system to image 3D ultrastructure using serial block-face imaging. Furthermore the system is equipped with the Zeiss Atlas V system to allow imaging of serial sections in a non-destructive manner.

  • FEI Tecnai 20 FEG with ICORR

The FEI Tecnai 20 FEG is a 200 kV TEM equipped with a Gatan 2001 energy filter with a 2k x 2K CCD camera, a Gatan US 4000 4k x 4K CCD camera and an FEI iCorr integrated light microscope for (cryo-) CLEM.

  • FEI T12 Spirit BioTwin

The FEI Tecnai 12 BioTwin is a 120 kV TEM with a LaB6 filament and a FEI Eagle 4k x 4K CCD camera to record high contrast images. The microscope has in-house developed software for automated scanning of specimen grids, and is equipped for cryo electron microscopy and tomography.

  • FEI T12 Spirit Twin with ILEM and STEM

The FEI Tecnai 12 Twin is a 120 kV TEM with a LaB6 filament and an FEI Eagle 4k x 4K CCD camera and is equipped with an integrated laser scanning microscope (iLEM) for correlative light-electron cryo microscopy and a Fischione HAADF STEM detector. Like the other TEMS this microscope runs in-house developed software for automated scanning of specimen grids and can perform cryo electron microscopy and tomography.

Sample preparation

Specimen preparation infrastructure includes instruments for carbon coating, plasma cleaning, high pressure freezing, freeze substation, (cryo-)ultramicrotomy and vitrification.

  • Emitech K950 Carbon coater and glow discharger
  • Emitech K975 carbon coater
  • Leica EM GP
  • Leica EM Pact 2 High Pressure Freezer
  • Leica EM AFS
  • Baltec CPD030 Critical Point Dryer
  • RMC ATUMtome
  • Leica EM UC6 microtome
  • Leica EM UC6 Cryo ultramicrotome
  • Leica EM UC7 ultracryomicrotome

Computing and visualization

Our computing infrastructure includes Windows and Linux workstations on which EM related software is implemented. We have dedicated hardware for GPU-based computations. We house two large multi-tile displays for the visualization of EM and LM datasets.