Cytometry : cell sorting & analysis

Flow cytometry measures the fluorescence or light diffraction of a large number of particles at high speed, such as cells, beads, bacteria, yeast, or organelles. At Institut Curie, flow cytometry is used mainly to quantify multiple markers on cells, with the option of simultaneously sorting multiple sub-populations of interest.

The primary advantage of flow cytometry is how quickly it produces data for a very large number of cells, allowing for complex and/or rare sub-populations of cells to be analyzed and sorted so that they can then be cultured or analyzed with molecular biology tools.

The cells in suspension may be simultaneously marked with 11 fluorochromes, each identifying a molecule of interest. The marked cells flow past a laser and, for each cell, the fluorescence intensity is quantified for each fluorochrome.
Fluorescent probes can be used to detect various parameters such as membrane potential, pH, or cell cycle (proliferation, apoptosis, etc).

The flow cytometry platform performs cell sorting, and also trains people to independently use high-speed multiparameter cytometers and the supporting software to acquire and analyze data.

Functions

  • cell sorting
  • training users on equipment and software
  • support for data acquisition and analysis
  • advice on cell preparation
  • results interpretation
  • support for preparing manuscripts, i.e. figures, documents, and methods

Institut Curie’s main flow cytometry facilities, which are shared between the Hospital and the Research Centre, are located in Laboratoire Laetitia at 26 Rue d’Ulm, Paris.

Equipment

  • 2 analyzers: one FACSort (4 colors) and one LSRII (11 colors)
  • 2 sorters: one FACSVantage (8 colors with UV) and one FACSAria (10 colors).

Contact and authorization to use the equipment

  • Analysis

The analyzers are self-service. Several other self-service cytometers are available at other Institut Curie locations, in both Paris and Orsay.
To use the self-service cytometers at the platform in Laboratoire Laetitia:
Register at the laboratory,

  • Call to schedule an appointment at +33 1 56 24 58 01 (office) or +33 1 56 24 58 02 (laboratory)

Training is required to use the self-service equipment without supervision. Theoretical training courses are given twice a year, and practical training is given once a month. All new users must contact the platform managers for their first independent session, or if they would like assistance from an engineer.

Sorting

Schedule an appointment with the managers:
Zofia.Maciorowski@curie.net ,  or

+33 1 56 24 58 01 (office) or +33 1 56 24 58 02 (laboratory).

Available in the Clinical Immunology unit (4th floor: 26 Rue d’Ulm):

  • One self-service AutoMACS magnetic bead-based cell sorter
  • One Luminex analyzer, for quantifying multiple analyses of very small volumes in a homogeneous phase, with an ELISA fluorescent bead system.