DONATE MAINS

cytometreThis platform provides to research teams powerful tools, expertise, technical assistance and technology monitoring with the aim at supporting research projects through high technology. It is available to the academic scientific community (e.g. CNRS, INSERM, Sorbonne University) as well as to the industry.

The staff is in charge of the proper functioning of the platform:

• organization of the activity (reservation planning, data storage)
• training / user support,
• project monitoring and analysis,
• maintenance and management of hardware and equipments.


Equipments

Cytomics FC500 Beckman Coulter

2 Lasers

- Argon ion laser, 488nm (20mW output)

- Helium - Neon ion laser, 633nm (20mW output)

Flow cytometry is a technique that allows high-speed analysis of a suspension of particles (cells, bacteria, parasites, beads, etc.). The individual characteristics of each of them such as the size, shape and complexity and any component or function that can be detected by a fluorescent compound. This cytometer does not allow cell sorting, however, depending on the applications, it may be carried out using magnetic beads.

Applications

  • Cell viability: 7AAD or PI staining
  • Apoptosis assays: annexin V-7AAD or PI, subG1 peak,
  • Caspase activity
  • Mitochondrial Membrane Potential, JC-1 staining
  • Cell proliferation, CFSE labeling, BrDU
  • Cell cycle using Wincycle software
  • Intracellular cytokines staining with or without monensin/brefeldin
  • GFP measurement
  • Antigen quantification in molecular equivalent soluble fluorochrome (MESF) or in antibody binding capacity (ABC)
  • Direct and indirect stainings, membrane or cytoplamic stainings
  • Cell characterization and phenotyping

Equipments

Bio-Plex MAGPIX Multiplex Reader Biorad

Luminex technology is based on the use of microspheres, colorful magnetic beads using a mixture of two fluorochromes whose ratio is known and clearly defined for each type of beads. There are different panels of 100 microbeads made according to the fluorochrome mixtures. These beads through their carboxylate groups, serve as solid support at different binding molecules covalently with amino groups. Luminex is a powerful multi-analytical system that can theoretically analyze up to 100 types of processes simultaneously in the same sample (25μL minimum).Several companies provide kits with beads coupled to the antibody, detecting one or more parameters (Biorad, Millipore, eBioscience, Life Technologies, etc.). The settings are specific (human, mouse, primate). Coupled beads "in way" may also be performed.

Applications

The different beads can be coupled to their surface with oligonucleotide probes, peptides or antibodies, thereby enabling the detection of alleles, antibodies or peptides


Team

FB newFrançoise Brignole-Baudouin
LR new
Luisa Riancho


Françoise Brignole-Baudouin
Scientific supervisor 

Email: fbrignoled@inserm.fr

Luisa Riancho
Technical manager

Email: luisa.riancho@inserm.fr