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Division of Nanoscopy Facilities

The division of Nanoscopy is equipped with cutting-edge scientific equipment.

M4I Facilities

Division of Nanoscopy Infrastructure

Cryo-electron microscopy is the only way to study cellular processes close to the in vivo situation. In order to do so, we have implemented a full workflow, starting with life cell imaging of cells or tissues followed by rapid cryo-fixation to preserve the structure. Cryo-light and Cryo-electron microscopy will resolve the 3D structure at nm resolution by means of electron tomography.

Our-full-cryo-correlative-workflow

Our full cryo-correlative workflow for ‘Selected volume tomography’

Step 1

The workflow starts on the CorrSight with live cell imaging of cell cultures on an EM carrier, which can be used throughout the complete workflow in order to prevent loss of orientation and sample contamination or destruction. Once a specific event or location has been identified by means of light microscopy and preserved by means of cryo-fixation, the vitrified sample can be re-examined on the CorrSight in a dedicated cryo-stage in order to precisely determine the position of interest.

Step 2

After transferring the sample to the Scios DualBeam, the identified region is thinned down to the appropriate thickness of about 200-300 nm without artifacts using the focused ion beam. The correlative MAPS software allows the transfer of all data from the CorrSight such as positions of interest and recorded images to the Scios DualBeam in order to know exactly where to prepare the lamella for subsequent high resolution cryo-TEM imaging.

Step 3

After thinning the samples to the proper thickness, the grids are loaded in the autoloader on the Tecnai Arctica for high resolution cryo-tomography. The Tecnai Arctica is equipped with FEI’s high sensitive Falcon direct electron detector for low-dose imaging, FEI tomography software for automated low-dose tomography and EPU software for automated single particle data acquisition.

More equipment

In addition to the CorrSight, Scios DualBeam, Tecnai Arctica, the Division of Nanoscopy also has the following equipment, which can be seen in the slideshow:

  • Ultramicrotome Leica EM UC7
  • Cryochamber Leica EM FC7
  • Leica EM AFS2
  • Combined iCorr fluorescence light microscope module and the FEI Tecnai electron microscope
Ultramicrotome-Leica-EM-UC7

Ultramicrotome Leica EM UC7

The Ultramicrotome Leica EM UC7 provides easy preparation of semi- and ultrathin sections as well as perfect, smooth surfaces of biological and industrial samples for TEM, SEM, AFM and LM examination.

Cryochamber-Leica-EM-FC7

Cryochamber Leica EM FC7

The Leica EM UC7 can be transformed into a Cryoultramicrotome by mounting the Cryochamber Leica EM FC7 to prepare cryo-sections ( -15° to -185°C) for TEM, SEM, AFM and LM.

Leica-EM-AFS2

The Leica EM AFS2

The Leica EM AFS2 performs freeze substitution and progressive lowering of temperature (PLT) techniques and allows low temperature embedding and polymerization of resins.

The Leica EM FSP (freeze substitution processor), an automatic reagent handling system combined with the Leica EM AFS2, dispenses reagents for both freeze substitution and PLT applications. The LED illumination from within the chamber and the attached stereomicroscope for viewing and positioning of samples ensures ease of use.

Combining-the-iCorr-fluorescence-light-microscope-module-and-the-FEI-Tecnai-electron-microscope

Combinie the iCorr fluorescence light microscope module and the FEI Tecnai electron microscope

Combining the iCorr fluorescence light microscope module and the FEI Tecnai electron microscope into a single, harmonized instrument affords users a streamlined approach to correlative microscopy. This workflow requires no sample transfer, allows capturing fluorescence data as well as high resolution electron microscopy data and correlates that data in minutes.

Watch the video for more information.

M4I office wing

The M4I office wing has been designed with the same open and transparent look and feel as our labs. Based on C.O.R.E. collaborative open research education. C.O.R.E. requires a transparent and open environment for both laboratories and offices. M4I has invested heavily in an innovative and open environment for collaborative research. Research and office space is shared by scientist from very different backgrounds and disciplines, ranging from the fundamental sciences, technology and engineering as well as clinicians. In line with the CORE philosophy of Maastricht University the infrastructure is primed for researchers to cross the boundaries of their own disciplines and stimulate each other to excel in translational imaging science.