Assistance in the design of preanalytical and analytical protocols for cytometers, assistance in sample handling, assistance in result exploitation, training on the principles of flow cytometry and on the use of cytometers based on user requirements.
Contacts
The team
- Unit manager: Laurent Poulain
- Research and training technician: Marilyne Guillamin
Activities
There is a great variety of research themes in the field; oncology, marine biology, toxicology, neuroscience and microbiology for example.
Samples are analysed direct with no specific markings for morphological analysis (size, granularity) and cell counts: algae population analysis, pollen sample analysis, fungal spore counts.
Other analyses are performed after using one or several specific markers such as intercalators or DNA-specific markers (cell cycle, ploidy, DNA replication, etc.) clusters of differentiation (CDs) (immunological analysis), retention or exclusion probes (viability and cell death), specific probes (free radicals, cell proliferation, transfection rate) or the use of marked beads (phagocytosis, cytokine study). The sensitivity of cytometry instruments also enables microparticles (in the region of 100 nm) to be studied.
After validating cytometry parameters with calibration beads, samples in suspension are analysed according to the desired parameters. Results are expressed in the form of mono- or bi-parameter histograms.
Technical resources
Gallios Cytometer Analyser (Beckman Coulter)
Equipped with 3 lasers (Blue: 488 nm; Red: 633 nm and Purple: 405 nm); a 32-tube carousel; detection of FS size (from 3 possible angles: 0.4 to 40 µm), of SS granularity and up to 10 fluorescences simultaneously (FL1 to 5 for the Blue laser, FL6 to 8 for the Red laser, FL9 and 10 for the Purple laser), analysed for 4 signal decades. Equipment acquired in 2011.
CytoFlex S Cytometer Analyser (Beckman Coulter)
Equipped with 4 lasers (Blue: 488 nm; Yellow: 561 nm; Purple: 405 nm and Near UV: 375 nm); detection of FS size, of SS granularity and up to 12 fluorescences simultaneously (2 on the Blue, 4 on the yellow, 4 on the Purple and 2 on the Near UV), analysis of microparticles (VSSC granularity on the purple, from 100 nm), analysis on 7 signal decades, direct measurement in absolute numbers (known sample volume), modular configuration. Material acquired in 2017.
Cytometry data analysis software
- CytExpert
- Kaluza
- FCS 6 Express
These software packages enable cytometry results to be presented in different manners (mono-or bi-parameter graphs, cell cycle analysis by phase, calculation of cell distribution in different generations, etc.)
Other laboratory equipment
- centrifuge (24 x 15ml tubes or 6 x 50ml tubes)
- micro-centrifuge (24 x 1ml or 2ml tubes)
- incubator
- PSM
- CellMacs Dissociator,
- programmable agitator with thermostat
- block thermostat (+4°C to +100 °C).
A few applications
Populations of populations: Micro-algae based on different fluorescences naturally emitted in a seawater sample (FL3 and 4) or pollen groups. (P2 mix)
Analysis of microparticles on the CytoFlexS with the V-SSC detector:
Immunological analysis of lymphocyte subpopulations.
Analysis of cell cycle and ploidy:
Anaylsis of micro-nuclei:
Analysis of division on different generations:
Partners
At regional level:
- SF ICORE, ANSES (Dozulé), Cyceron, LARIA, etc. research teams
At national level:
- C-Ris Pharma (35)