The flow cytometry platform offers several services (multi-color analysis, single and population cell sorting, etc,) as well as developing new techniques as required by the laboratories on the campus.

The used techniques are flow cytometry, imaging, sorting using antibodies coupled to magnetic beads.

Functioning Our activity is essentially based on flow cytometry analyses, image analysis and cell sorting. For analysis, machines are ‘self-service’ and available on reservation basis. Adequate training is mandatory before having opened access to the analytic flow cytometers, and varies according to the complexity of the analysis required (fluorescent parameters, etc). Support for data interpretation is also provided. A preliminary interview enables us to establish with our “customers” the protocol for preparing the cells and to plan the project.

Objectives:

To create a productive exchange with PFC users in order to develop new tools, techniques and approaches relevant to analytical and preparative flow cytometry.

To improve our efficiency and therefore availability for sorting (throughput).

To develop collaborations with our tools’ providers to meet the needs for the campus (development of the expertise on pathogenic hardware, sorting after analyzing 6 colours with 4 ways, complex analyses on more than 9 colours...)

To work with support services in Pasteur Institute like “Hygiene & Security”, “Quality”, “Health at Work” and “Logistics” for improving our performance.

Conclusions: Forty research teams (for the most part from the Pasteur Institute) are the major users of the PFC. Annual services include approximately 5000 analyses and 300 sorts. Experiments usually involve the detection of membrane antigens (between 3 and 9 fluorochromes measured simultaneously, according to the apparatus used), but also include the measurement of calcium flux, cell cycle, FRET and apoptosis assays.

The platform proposes hands-on trainings regularly each month. Every year, about 180 trainings have been organized.

The team interacts with internal and external resources for improving efficiency: to identify new technological instrument, to bring appropriate resolution to the Campus’ problems…

Prospect: The PF, which provides a great number of service, evolves with its human resources to meet the researchers’ needs in compliance with the Pasteur’s scientific politics by:

1. Identifying and testing new tags.

2. Developing new methodologies implying the cytometry.

3. Increasing complex expertise and sorting of pathogenic samples. The team is working on sorting and analyzing bacteria and viruses.

4. Evolving new forms of training courses, adapted to the software recently acquired (for controlling the new cytometers) by the Laboratory

5. Improving: Quality for validating the rough data; H&S for securing expertise on pathogenic samples and evolving our risk’s management; Logistic for making smoother the use of the core-facility.controlling the new cytometers) by the Laboratory.

1. Improving: Quality for validating the rough data; H&S for securing expertise on pathogenic samples and evolving our risk’s management; Logistic for making smoother the use of the core-facility.

Imagopole which includes the PFCF has been just certified ISO 9001, version 2000 by AFAQ (Organism of Quality).

Keywords: fluorescence, cytometry, cell analysis, cell sorting, services

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Similar functional activity of dendritic cells recruited to the mesenteric lymph nodes of newborn and adult mice after the rectal delivery of Mycobacterium bovis BCG. Microbes and Infection 8 (9-10) ; 2341-51. PMID: 16872860

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