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Flow cytometry

 

The platform offers technical expertise and state-of-the-art equipment for flow cytometry analysis and sorting.
Our offerings include support for experimental design, sample acquisition, data analysis, as well as providing cell sorting services.

Principle

Flow cytometry is a high-throughput technique that enables multiparametric and quantitative analysis at the single-cell level.

The suspension of cells (or elements) is drawn into a quartz flow cell by a pressure system (or peristaltic pump). The cells are then centered in a narrow liquid vein by a sheath liquid (hydrodynamic focusing). The cells (or elements) are illuminated sequentially by one or more laser beams. The signals emitted and of interest (scattering properties, autofluorescence, fluorescence) are collected by photodetectors (photomultiplier tube or photodiode). The light signal is converted into an electrical pulse, then into an analog signal.

Several signals are collected:

  • (Forward Scatter, FSC), which provides information on particle size,
  • Side Scatter (SSC), which provides information on the internal structure of the cell,
  • Fluorescence emission

Expertise

  • Immunostaining
  • Phenotyping
  • Cell cycle, proliferation assay, genome size
  • Viability, Apoptosis
  • Expression of fluorescent reporter protein (GFP, mCherry, mScarlet, FRET sensor…)
  • Functional analysis, metabolic flux
  • Morphology analysis
  • Analysis of rare events
  • Fluorescence-activated nuclei sorting (FANS)

Our equipment

Analyzer: Attune CytPix (ThermoFisher)

Optical configuration:

  • Blue: 488nm (3 parameters)
  • Red: 640nm (3 parameters)
  • Violet: 405nm (4 parameters)
  • YG: 561nm (4 parameters)

High-speed brightfield camera

Sorter: FACSAria Fusion (BD)

BSL2
Nozzles: 70µm; 85µm; 100µm; 130 µm
1-4 way sorting channels: 1.5 mL tubes; 2mL; 5mL; 15mL; 6, 12, 24, 48, 96, 384-well plates; slides; custom

Optical configuration:

  • Blue: 488nm (4 parameters)
  • Red: 633nm (3 parameters)
  • Violet: 405nm (4 parameters)
  • YG: 561nm (4 parameters)
  • UV: 355nm (3 parameters)

Data analysis

Various analysis software packages are available: FlowJo, Diva, FCS Express, Attune software.