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  Director : S.L.SHORTE (sshorte@pasteur.fr)


  abstract

 

In its capacity as a core facility, established in 2001, the CID is a well equipped "technical-platform" PT-unit providing expertise and hardware to aide researchers in designing, performing and analysing experiments using digital imaging microscopy. In particular, our focus is on dynamic imaging in living cells and tissues in order to gain insight on questions concerning cell biology, cell signalling, cell/tissue organisation and infectious processes therein. The facility is open to researchers inside and outside Pasteur, who benefit both from the wide variety of cutting-edge techniques offered, and our commitment to pursuing challenging projects requiring novel solutions through collaborative R&D efforts.



  report

cale

The Centre d'Imagerie Dynamique (CID) is a new technological platform at the Institut Pasteur comprising a part of the Plateaux Techniques (PT) initiative instigated by the Direction des Equipments et Technologies Stratégiques (DETS). Specifically, the CID mission is to provide and develop high-level cellular imaging technology and expertise for addressing biological questions, which require visualisation and measurement of dynamic processes inside living cells. Albeit primarily a technical platform open to all, the CID is more specifically integrated into the Pasteur scientific community through a direct affiliation with the department of Cell Biology & Infection (P.Sansonetti), and has strong links to other departments including Developmental Biology (M.Buckingham), and Molecular Medicine (J.L.Virelizier). The combination of technical and scientific research-engineering expertise inside the CID renders this PT-unit a resource, affording Pasteur's researchers a unique and dedicated scientific service.

During July 2002, the equipment comprising the PT-Centre d'Imagerie Dynamique (CID) was installed into its new permanent location, a purpose-built laboratory facility comprising 10 independent imaging suites plus space for cell-tissue preparation and informatics. The purpose designed P2-level imaging facility comprises an ergonomic 200m2 "Biological Laboratory" environment (Bât.Monod) allowing imaging experiments on living tissue to be performed entirely inside the facilities. In addition, during 2002, we realised the installation of a high-speed 4-D imaging system in a P3 laboratory, in collaboration with the Dept. Molecular Medicine. For all systems, following appropriate training in the use of the microscopes and other equipment, our goal is to enable researchers with autonomous access to the facilities, where they can operate in a "home-from-home" scientific environment.

Complementing our service function, the CID also harbors and facilitates local, national and international collaboration initiatives involving biological imaging. For example, the "European Light Microscopy Initiative" (ELMI), through our partnership in the European funded EAMNET (European Advanced Microscopy Network), and in this capacity the CID participated in hosting the ELMI annual meeting (May 2002). Equally important, at a local level, the CID is also committed to support specific internal research programmes and in particular the "PTR" and "GPH" initiatives. Finally, the CID provides for specialised teaching/training courses for microscopy, and during 2002 (in collaboration with Zeiss, France) launched the first in a series of AIM (Advanced Imaging Microscopy) training-workshop courses that was attended by more than 50 people. During 2003 further courses are planned, and will be announced shortly.

Fuelling the above activities the CID puts a strong focus upon development and realisation of novel and advanced imaging techniques. Importantly, these initiatives are carefully tuned to the institute's research needs, in a grass-roots manner, involving continuous discussion with groups across the campus. During 2002 several projects have been developed and have proven their utility. Furthermore, one has yielded a European patent. Current topics of interest include:

-Three-dimensional visualisation of spatial signals inside non-adherent living cells

-Bioluminescence to measure biological activities inside single living cells, and/or tissues

-High-Speed 4-D microscopy

Equipment. The CID currently maintains twelve "state-of-the-art" microscopes, each customised for specific applications. Off these two are outside the central facility, including one high-speed 4-D "TILL" system placed in a P3 laboratory.

Single Photon Confocal Microscopes:

Zeiss LSM510 — Ar/He/Ne laser, equipped for FCS, FRAP, FRET and 3-/4-D imaging

Zeiss LSM510 — Ar/He/Ne laser, equipped for 3-D imaging

Leica TCS-4D Kr/Ar laser (permenantly situated in Lwoff building)

Perkin-Elmer — Nipkow spinning disk system for fast confocal imaging in live cells.

Two-Photon Microscope.

Zeiss LSM510 For "deep" in vivo and in situ4-D tissue visualisation.

Epi-fluorescence Digital Imaging Stations.

Three TILL "Fast" Imaging Systems equipped for quasi-real-time multi-dimensional imaging

Three Zeiss Multi-dimensional fluorescence time-lapse imaging systems.

One Quantitative "real-time" bioluminescence/fluorescence system enabling measurement of bioluminescence

from single living cells using promoter driven luciferase, or colentrazine,

IRIX based Hardware/Software:

Two SGI Octane2 Workstations plus 1 SGI ORIGEN 3400 Server (IRIX 6.5)

Huygens/BitPlane Image processing software for 3-4-D reconstruction/deconvolution.

Microinformatics- 5 "high-end" PC's running various image analysis/processing and visualisation software for customised image processing/enhancement, morphological, numerical, algorithmic, and counting operations.

Cell preparation and maintenance

  • P2 cell-culture room

  • Tissue preparation room

  • 3 Bioptechs Live Cell Incubation Chambers

  • Single cell microinjection

  • Evotek microscope based single cell sorting, and 3-D manipulation.

Activity turnover.

Currently, we host around 75 experimental sessions (each lasting 3-6hrs) per month.

Techniques under development.

  • Pulsed-"flash"-laser photolysis, and photo-activation

  • Intra-vital Bioluminescence imaging.

  • Confocal Endoscopy

Patents accepted.

"Method and device for 3 dimensional imaging of suspended micro-objects providing high-resolution microscopy" (EP 02 292 658.8, 25 October 2002).

Projects under development.

  • Quantitative "real-time" bioluminescence measurement of gene expression in single living cells using promoter driven luciferase, or colentrazine,

Posts available

We have ongoing opportunities for motivated individuals with training in imaging, informatics, physics and/or biology. Contact us: cid@pasteur.fr

Keywords: Digital imaging, Cell Biology and Infection, Confocal and 2-Photon Microscopy, Image Analysis, Fluorescence



  web site

puce More informations on our web site


  publications

puce Publications of the unit on Pasteur's references database


  personnel

  Office staff Researchers Scientific trainees Other personnel
        PERRET, Emmanuelle, (Technicien,eperret@pasteur.fr)

ROUX, Pascal, (Engineer,proux@pasteur.fr)

SHORTE, Spencer (PhD, Engineer,sshorte@pasteur.fr)


Activity Reports 2002 - Institut Pasteur
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