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Flow cytometry

Flow cytometry is a technique to analyze the physical and functional characteristics of each cell within a suspension of cells. This method is used to determine the proportions of different cell subpopulations in a cell culture. It can also measure a large number of cells very quickly for the parameters of cell size and DNA content as well as the presence of specific surface, intracellular or intranuclear molecules. These molecules are detected with fluorescent probes and lasers.

Today, new generations of devices are equipped with up to 7 lasers that measure over 30 different variables. Some devices sort cells according to these parameters and isolate them into cell subpopulations for further analysis or culture so that their functions can be studied.

Individual training on each system is available by appointment. The BD FACSAria III cell sorter has a negative pressure hood to sort potentially infectious samples, including human cells from healthy donors.

Equipment

BD LSRFortessa X-20

  • The BD LSRFortessa X-20 has 4 lasers that detect 16 fluorescence parameters in addition to cell size (FSC) and cell structure (SSC) parameters.

    • Violet (407 nm): 6 fluorescence detectors
    • Blue (488 nm): 3 detectors (SSC and 2 fluorescence detectors)
    • Yellow-green (561 nm): 5 fluorescence detectors
    • Red (633 nm): 3 fluorescence detectors

BD LSR II

  • The BD LSR II has 4 lasers that detect 10 fluorescence parameters in addition to cell size (FSC) and cell structure (SSC) parameters.

    • Blue (488 nm): 5 detectors (SSC and 4 fluorescence detectors)
    • Red (633 nm): 2 fluorescence detectors
    • Violet (407 nm): 2 fluorescence detectors
    • UV (355 nm): 2 fluorescence detectors

    The filters and mirrors on this instrument can also be changed to detect different dye combinations.

BD FACSCanto

  • The CANTO A has 2 lasers that detect 6 fluorescence parameters in addition to cell size (FSC) and cell structure (SSC) parameters.

    • 488 nm: 5 detectors (SSC and 4 fluorescence detectors)
    • 633 nm: 2 fluorescence detectors

2 BD FACSCalibur

  • The Caliburs have a blue and red-diode laser to detect 4 fluorescence parameters in addition to cell size (FSC) and cell structure (SSC) parameters.

    • 488 nm: 4 detectors (SSC and 3 fluorescence detectors)
    • 633 nm: 1 fluorescence detector

BD ARIA III Sorter

  • The BD ARIA III has 4 lasers that detect 16 fluorescence parameters in addition to cell size (FSC) and cell structure (SSC) parameters.

    • Violet (407 nm): 6 fluorescence detectors
    • Blue (488 nm): 3 detectors (SSC and 2 fluorescence detectors)
    • Yellow-Green (561 nm): 5 fluorescence detectors
    • Red (633 nm): 3 fluorescence detectors

    Nozzles of different sizes (70, 85 and 100 µm) and various differential pressures are used depending on the type of cells to be sorted. Four populations can be sorted simultaneously, in addition to the plate sorting function.

SONY SH800 Sorter

  • This SONY Sorter has two collinear lasers (488 nm and 561 nm) that detect 6 fluorescence parameters in addition to cell size (BSC) and cell structure (SSC) parameters.

    The system offers 70 µm and 100 µm chips, and the pressure can be changed for the type of cells to be sorted. The SONY can sort 2 populations simultaneously and has a plate sorting function.

    Software
    Through the cytometry platform, researchers have access to BD CellQuest Pro, BD FACSDiVa, BD FCAP Array, ModFit (Verity Software), FlowJo, FlowLogic and Cytobank.

Coordinator

Martine Dupuis, MSc
514-252-3400, extension 3521