Electron microscopy facility

The Electron Microscopy Facility (EMF) of the Institut de Biologie Paris-Seine offers a set of skills and equipment in transmission and scanning electron microscopy to the scientific community (IBPS, UPMC, public and private laboratories). A team of four engineers animate the EMS. They provide methodological support, user training, maintenance of the equipment and scientific activities (seminars, teaching).

EMS provides access to a wide range of equipment allowing the preparation and the observation of a large variety of biological models, from bulk specimens or cell cultures to thin films or nanoparticles. Sample preparation dedicated to ultrastructural or immunolocalisations studies can be realised "conventionally", with chemical methods or using cryomethods. The facility has a long-lasting expertise in cryomethods, which associate a fast immobilisation of samples with an optimal preservation of their integrity (high-pressure freezing, freeze substitution, plunge-freezing, freeze-fracturing, freeze-etching).

Our park is equipped with a Scanning Electron Microscope (SEM) to visualise the sample surface. Furthermore it includes two Transmission Electron Microscopes (TEM) to observe standard sections of samples (80-120 kV), or to perform tomographic or cryo-TEM analysis (200 kV).

The service is also involved in various technological developments to provide innovative methodological solutions, for example: optimal sample preservation (cryomethods) 3D reconstruction (energy-filtered imaging tomography) or screening for rare events detected with fluorescence (correlative microscopy).

Material

  • Transmission Electron Microscope 80-120kV (912 Omega Zeiss) + side-mounted digital camera for large field of view acquisitions (Veleta, Olympus)
  • Transmission Electron Microscope 200kV (2100HC, JEOL) + post-column energy filter (Gif-Tridiem, Gatan)
  • Dock-station and sample holder for cryo-microscopy (Gatan) + high-tilt sample holder for electron tomography (JEOL)
  • Scanning Electron Microscope equipped with a tungsten filament (Stereoscan 260, Cambridge) + digital acquisition system
  • Vibratom (VT1000S, Leica)
  • Ultramicrotomes (UCT, Leica et Ultracut E, Reichert-Jung)
  • Plunge-freezing apparatus (CPC, Leica)
  • High pressure freezing (HPM100, Leica)
  • Freeze-substitution apparatus (AFS, Leica)
  • Freeze-fracturing and freeze-etching apparatus (BAF 400T, Balzers)
  • Critical point drying apparatus (CPD7501, Quorum)
  • Gold sputter coater (Scancoat Six, Edwards)
  • Data treatment and tomographic reconstruction (Imod, TomoJ)

Services

Presentation of offers:

EMS is mainly opened to biology laboratories of the Institute. All projects developed within the platform may include a conception monitoring[iS1], follow-up, realisation and result analysis. An interview precedes each project to evaluate feasibility and to propose the most appropriate techniques.

Type of offers:

  • Sample preparation in TEM and SEM
  • Sample preparation and image acquisition
  • Observation sessions for autonomous users
  • Engineer-assisted observation sessions
  • Long-term collaborative projects

Methodologies

  • Conventional sample preparation and observation in TEM: cytochemistry ultrastructurale, immunocytochemistry, negative contrast
  • Ultramicrotomy
  • High pressure freezing, freeze-substitution
  • Freeze-fracturing and freeze-etching
  • Cryo-microscopy in TEM
  • Tomography and cryo-electron tomography
  • Correlative light electron microscopy
  • Conventional sample preparation and observation in SEM

Methodological development

The electron microscopy facility introduced spectroscopy and energy-filtered imaging in TEMA, allowing chemical microanalysis on biological samplesB,C and three-dimensional reconstructions on thick sections in tomography and cryo-tomographyC. The facility has also pioneered the introduction of cryomethods in France in late 90's to develop optimal preservation of biological samples.

As of January 2016, our platform will house a high-resolution SEM coupled to cryopreparation and automatic imaging of serial sections. This will permit high-resolution viewing of cells and tissues maintained in their native state, at the subcellular and molecular levels, and high contrast TEM-like imaging, with a very wide field of view and 3D visualization of large volumes.

AShillito B., 1997, J Struct Biol. Oct;120(1):85-92.

BLechaire JP., 2006. Biol Cell. Mar;98(3):163-70.

CBoudier T., 2005, J Struct Biol. Aug;151(2):151-9.