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     Cytometry (Platform)


  Director : Anne-Marie Balazuc (abalazuc@pasteur.fr)


  abstract

 

The platform of cytometry proposes prestations of services, for analyses and sorting of cells.

The techniques used are the flow cytometry, cytometry in image, microscopy confocale, the sorting using antibodies coupled to magnetic balls. It also takes part in the development and the optimization of these techniques, like with the training of the interested teams.



  report

cale

The flow cytometry core-facility is integrated into Imagopole

Current organization

MCW: Responsible of the pole of analyzes by cytometry in flow.

She ensures the follow-up methodological and technological analyzers, develops the formations with acquisition-analyzes according to the evolution of the park of the cytometers and the needs . She collaborates with the users who wish to profit from her expertise either to interpret their results, or to develop their analyses.

AL: Responsible of the pole of cellular sorting.

She carries out expertise on the MOFLO (DAKOCytomation), screening machine high performance and ensures the training the users of the autoMACS for cellular sorting by magnetic balls. She also ensures the methodological and technological follow-up screening machine in relation to the providers (qualification of the equipment) and the users (development of the expertises).

HKB: person in charge for the development of expertise on pathogenic samples de type P2 & P3*.

Expert 1/ in cytometry in image: she does analyses with the ACAS; 2/ in Cytometry in flow: she carries out expertise on the ARIA (the new screening machine high performance which has been just acquired by the platform). She ensures the maintenance of the autoMACS and the theoretical part of the formation the analysis.

AMB: Responsible of the platform.

She frames the team and mobilizes it towards the objectives, ensures the management of financial and human means, organizes and plans the activities of the platform. She also ensures the communication of the laboratory on the services and attends the meetings of the committee users. She manages the resolution of problem by the means of the Qualité step which she initiated on the PFC. She takes care of the organization of the expertises on pathogenic samples (acquisition of new screening machine FACS Aria and enclosure PSM of the type II).

Our activity is essentially based on analyses (in flow or image) and sorting of cells, in the form of "provisions of services" at the request of the research teams. A preliminary maintenance enables us to stop with our "customers" the protocol of preparation of the cells and to plan handling. We help them if necessary to interpret their results...

According to the complexity of the apparatus, this one is handled by our expert or placed at the disposal of the users in "self-service". In this last case, an obligatory formation is organized beforehand by the PF.

Objectives.

To animate and manage the group dynamics initiated with the users of the PTF.

To improve our organization to gain in effectiveness and of availability.

To develop in collaboration with the providers of the tools to meet the needs for the campus (developed of the expertise on pathogenic hardware, sorting 6 colors with 4 ways, complex analyses on more than 9 colors...)

To be integrated within Imagopole: for better defining our functions on the level scientific, technical, of engineering and service...

Conclusion

Our equipment (a cytometer in image with laser scanning, of 3 sorting machines of cells, 5 analyzers, a microscope confocal) makes it possible in particular to carry out sorting at high speed, by separating 4 pennies cellular populations simultaneously. The analyzers are equipped in a complementary way making it possible to measure fluorescent signals with excitation laser UV 325 nm, violet 405 nm, bleu 488 nm et rouge 630 nm.

Forty teams, exits very mainly of the Pasteur Institute profit from these services (5000 analyses and 300 sorting approximately). The experiments relate mainly to the detection of membrane antibodies (between 3 and 7 fluorochromes measured simultaneously, according to the apparatus used), the measurement of calcic flows, the study of the cellular cycle, FRET and the apoptose.

The PF, which provides a great number of service, evolves/moves with the measurement of its human resources to meet the needs for the researchers:

By identifying and testing new fluorochromes (for example Pacific blue, Cascade yellow, BFP and CFP).

The introduction or the development of new methodologies implying the cytometry.

Increasingly complex Tri (6 colors, 4 ways) and sorting of pathogenic samples.

The platform proposes hands-on trainings regularly each month, from which this year 100 people profited. New forms of training courses, adapted to the hardware recently acquired by the Laboratory are regularly developed.

Keywords: fluorescence, cytometrie, microscopy confocale, analyze cells, sorting of cells



  web site

puce More informations on our web site


  publications

puce Publications 2005 of the unit on Pasteur's references database


  personnel

  Office staff Researchers Scientific trainees Other personnel
        KIEFER BIASIZZO Hélène, technicienne kiefer@pasteur.fr

LOUISE Anne , ingenieur alouise@pasteur.fr

WAGNER Marie Christine, technicienne, mcwagner@pasteur.fr


Activity Reports 2005 - Institut Pasteur
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