Help with the design of pre-analytical and analytical protocols on cytometers, assistance with sample runs, help with processing results, training on the principle of flow cytometry and on the use of cytometers according to requirements.
Contact
The team
- Manager: Laurent POULAIN
- Research and training technician: Marilyne GUILLAMIN
Activities
The research fields involved are very varied: cancerology, marine biology, toxicology, neuroscience and microbiology, for example.
Samples are analyzed directly without specific markings for morphological analyses (size, granulosity) and cell counts: algae population analysis, pollen sample analysis, fungal spore counting.
Other analyses are carried out using one or more specific markers, such as intercalants or DNA-specific markers (cell cycle, ploidy, DNA replication, etc.), CDs differentiation clusters (immunological analyses), retention or exclusion probes (viability and cell death), specific probes (free radicals, cell proliferation, transfection rate) or the use of labelled beads (phagocytosis, cytokine studies). The sensitivity of cytometry equipment also enables analysis of microparticles (around 100 nm).
After checking cytometer parameters with calibration beads, suspended samples are analyzed according to the desired parameters. Results are expressed as mono- or bi-parametric histograms.
Technical resources
Gallios Analyzer Cytometer (Beckman Coulter)
Equipped with 3 lasers (Blue: 488 nm; Red: 633 nm and Violet: 405 nm); 32-tube carousel; detection of FS size (at 3 possible angles: 0.4 to 40 µm), SS granulosity 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 Violet laser), 4-decade signal analysis.
Equipment acquired in 2011.
Cytometer Analyzer CytoFlex S (Beckman Coulter)
Equipped with 4 lasers (Blue: 488 nm; Yellow: 561 nm; Violet: 405 nm and Near UV: 375 nm); detection of FS size, SS granulosity and 12 fluorescences simultaneously (2 on Blue, 4 on Yellow, 4 on Violet and 2 on Near UV), microparticle analysis (VSSC granulosity on Violet; from 100 nm), 7-decade signal analysis, direct measurement in absolute value (sample volume known), modular configuration.
Equipment acquired in 2017.
Cytometry data re-analysis software
- CytExpert,
- Kaluza,
- FCS 6 Express.
These software packages can be used to present cytometry results in a variety of ways (single- or bi-parametric graphs, cell cycle analysis by phase, calculation of cell distribution in various generations, etc.).
Other laboratory equipment
- a centrifuge (24 tubes 15ml or 6 tubes 50ml),
- a micro-centrifuge (24 x 1ml or 2ml tubes),
- an oven,
- an SHP,
- a CellMacs Dissociator,
- a thermostatically-controlled programmable agitator,
- a thermostatic block (+4°C to +100°C).
Some applications
Populations: Microalgae based on differences in naturally emitted fluorescence in a seawater sample (FL3 and 4) or pollen set. (P2 mixture)
Microparticle analysis on the CytoFlex S with the V-SSC detector :
Immunological analysis of lymphocyte subpopulations :
Cell cycle and ploidy analysis :
Microkernel analysis :
Division analysis across generations :
Partners
At regional level :
- SF research teams ICORE, ANSES (Dozulé), Cyceron, LARIA...
At national level :
- C-Ris Pharma (35)