RIO Imaging             Iso 9001

The imaging platform of the Institute for Neurosciences of Montpellier is part of the Regional Imaging Platform (RIO) of Montpellier since the creation in 2004 (MRI: www.mri.cnrs.fr). It brings together scientific and technical knowledge dedicated to Neuroscience Imaging. In addition, the platform features various High Technologies in Microscopy for fixed or alive samples, and image analysis solutions. The platform is certified ISO 9001:2008 (LRQA) and labeled IBISA (Biology, Health and Agronomy Infrastructures: www.ibisa.net). It is accessible to public or private external laboratories.

We have different missions:

  • Providing an access to analysis equipment
  • Assisting in the preparation and observation of samples
  • Adapting and developing new in vitro and in vivo applications.
  • Automating image processing
  • Expertise
  • Training and Teaching.

1. MULTI-PHOTON MICROSCOPY

Multi-photon microscopy is a powerful tool for imaging deep tissues and has many advantagesover conventional single-photon microscopy for biological applications. The large separationin wavelength between excitation and fluorescence provides a good signal to noise ratiowithout sacrificing the fluorescence signal. It is a powerful technique for 3D imaging becausethe biological materials are transparent to infrared frequencies and can be excited to penetration depths inaccessible to one photon confocal microscopy. This spatial localization of excitation also allows a significant reduction of photobleaching static cells, biological molecules and fluorophores. In addition to the fluorescence signals produced by the process of multiphoton excitation, nonlinear phenomena, such as second harmonic generation (SHG) provide useful information on the structure and optical properties of a specimen.

Zeiss LSM 7MP / OPO


ZeissLSM7MP OPO

Multi-photon microscope Zeiss upright AxioExaminer. Equipped with a multi-photon laser Ultra II Coherent Chameleon (wavelength range between 680 and 1080nm) coupled to a Compact Chameleon OPO (Optical Parametric Oscillator) Coherent (wavelength range between 1000 and 1300nm System).

 

    • Imaging in vivo multi-dimensional, multi-color
    • Imaging on small animals
    • Fluorescence, GHS, TGH
    • Environmental chamber
    • Detection: 2PMT classic NDD and 2 BiG PMT with photon counting possible
    • Microscope: Motorized Zeiss AxioExaminer upright microscope


2. CONFOCAL MICROSCOPY

Confocal microscopy is an optical imaging technique used to increase optical resolution and contrast of a micrograph by using point illumination and a spatial pinhole to eliminate out-of-focus light in specimens that are thicker than the focal plane. It enables the reconstruction of 3D structures from the obtained images. This technique has gained popularity in the scientific and industrial communities and typical applications are in life sciences, for some reasons: high-speed imaging, reduced photobleaching. Spectral imaging can be done for a signal analysis in the frequency domain.

Confocal 1 Zeiss LSM 700  

Confocal1ZeissLSM 700

LSM 700 confocal laser scanning (point-scaning) with a single pihole. A secondary dichroic (VSD: Variable Secondary Dichroic 400 to 700nm), separates light emission between two detectors (PMT). It is led by the Zen software.

 

 

 

 

  • Samples fixed
  • Multi-dimensional imaging, multi-color
  • Excitation: 488nm laser diode, 555nm and 633nm
  • Detection: 2 PMT for fluorescence
  • Microscope: Motorized Zeiss Axio Observer Z1 inverted microscope


Confocal 2 Zeiss LSM 5 Live DUO  

Confocal2ZeissLSM 5Live DUO
Confocal Zeiss 5 Live Duo with two scan heads: LSM 510 point-scanner, the conventional relatively slow to allow imaging of the multidimensional fixed tissues and LSM 5 Live line-scanner that has maximum speed (120 frames/s with 512x512 pixels). It is a suitable tool for the observation of rapid dynamic processes on sensitive live preparations, such as nerve cells.


    • Environmental chamber for live cells
    • High-speed acquisition
    • Low-light live imaging
    • Spectral imaging
    • Automated multi-position XYZT acquisition
    • Z-sectioning and 3D rendering
    • Excitation: laser lines at 405, 458, 477, 488, 514, 545, 561, 633 nm.
    • Microscope : Motorized Zeiss Axio Observer Z1 inverted microscope


Confocal 3 Leica SP5 X  

Confocal 3 Leica SP5 X

Leica SP5 confocal with a Supercontinuum Laser White - AOBS - spectral detectors. Allows you to select specific wavelength excitation with nanometer precision, the choice of the emission band collected, the ability to excite up to 8 lines simultaneously without changing dichroic.

 

    • Live and fixed samples
    • Spectral imaging excitation and emission
    • Z-sectioning and 3D rendering
    • AOBS based system
    • Excitation: 405nm laser diodes, Supercontinuum Laser (470nm-670nm)
    • Detection: 3 PMT for fluorescence and one transmission
    • Microscope: DMI6000


3. CONVENTIONAL (WIDEFIELD) MICROSCOPES

Widefield 1 - Zeiss Axioimager Z1 / Zen

Widefield 1


Zeiss AxioImager microscope (Z1 upright) fully motorized and equipped with an Apotome module and a Axiocam camera MRm. Dedicated to image acquisition of fixed samples as well as in fluorescence light background. The Apotome module enables optical sections through fluorescently labeled biological samples quickly.

 

    • Samples fixed
    • Module Apotome (Structured Illumination)
    • Automated Stage
    • Brightfield, DIC, Multicolour Fluorescence
    • Zen Software

Widefield 2 - Zeiss Axioimager D1 / Axiovision

Widefield 2



Zeiss microscope right AxioImager D1 with a color CCD camera AxioCam MRc. Dedicated to image acquisition of fixed samples as well as in fluorescence or Brightfield. AxioCam MRc color CCD camera is perfect for histological stains.

 

 

    • Samples fixed
    • Brightfield, DIC, Multicolour Fluorescence
    • AxioVision Software

Widefield 3 - Zeiss Axiovert 200M / Metamorph

Widefield 3
Zeiss Axiovert inverted 200 M microscope fully automated equipped with a controlled atmosphere chamber, a motorized stage and a camera (Roper Micromax). Controlled by the MetaMorph (Universal Imaging) software, used to monitor for several days cell cultures in boxes of 6 and 12 wells, white light and fluorescence due to 1-122 in separate fields or adjacent wells (mosaic image).

    • Living Samples
    • Environmental chamber for live cells (37 ° C 5% CO2)
    • phase contrast, DIC, Fluorescence

Widefield 4 - Zeiss Axiovert 200 / Metafluor

Widefield 4

Station developed based on a Zeiss Axiovert 200 inverted microscope, a changer fast wave length (Sutter Lambda DG-4) and a camera Coolsnap ES. The system, driven by the MetaFluor (Universal Imaging) software allows to observe the response of many neurons at the same time with a frequency of 3-4 frames per second.

 

    • Living Samples
    • Ratiometric imaging (calcium imaging…)
    • Perfusion medium
    • Phase contrast, DIC, Fluorescence


4. DIGITAL SLIDE SCANNER / NANOZOOMER

DigitalSlideScanner
Imaging systems to high capacity, a robot dedicated to immunohistological labeling (color image). Essential for toxicity studies, high throughput screening and storage of virtual slides after staining or immunostaining tools.

 

    • Automatic Acquisition (210 slides)
    • Histology staining (color images)
    • Fluorescence
    • Provision of service for digitizing histological slides in brightfield or fluorescence.


 5. MACROSCOPY

ZeissDiscoveryV20

Zeiss Discovery V20 Stereo zoom microscope with transmitted, reflection and fluorescence illumination, 1x and 2.3x lenses, filter sets for DAPI, GFP and DsRed and a AxioCam MRm CCD camera.

 

 

    • Macroscopic & microscopic imaging
    • Motorized 20:1 zoom
    • Illumination and contrast methods based on cold light or LED
    • LED transmitted illumination
    • LED reflected brightfield illumination
    • Axiovision software


6. IMAGE ANALYSIS AND DECONVOLUTION WORKSTATIONS

Two workstations with image analysis software:

    • Imaris (Bitplane) & Volocity (Perkin Elmer) for multidimensional data analysis
    • Huygens (SVI) for deconvolution of image stacks
    • Zen & Axiovision for Ziess files
    • MetaMorph offline (Tracking, Count nuclei, neurite outgrowth…)
    • Image J