Flow Cytometry Facility

The selection and purification of molecules, cells, tissues, and organisms of interest are critical yet often time-consuming aspects of biomedical research. Flow cytometers have revolutionized the sorting and analysis of large numbers of individual cells, microorganisms, and other particles at high speed. The fluidics subsystem of a flow cytometer brings particles in suspension into the flow cell where they are excited by light sources of different wavelengths. With the use of multiple mirrors, band pass, and long pass filters, the emitted light is routed to the appropriate photomultipliers, where the incoming photons are converted into electrons and visualized for further analysis. Applications range from simple cellular readouts to complex phenotyping and high-throughput screening.

Our facility houses three different machines that support a broad range of applications:

BD FACSymphony A1 cell analyzer with small particle detector:

The BD FACSymphony A1 is used for analyzing particles, ranging from very small ones such as extracellular vesicles, to microorganisms such as yeasts, and a wide variety of blood and tissue cells. This analyzer supports high-resolution analysis of up to 16 parameters simultaneously. Examples of possible applications are measuring cell proliferation, apoptosis, transfection efficiency, cell cycle progression, cytometry bead array, and identification of rare subsets by multicolor staining (both intra- and extracellular).

BD FACSAria Fusion cell sorter:

The BD FACSAria Fusion is used for sorting particles, ranging from microorganisms such as yeasts to various blood or tissue cells. This is performed at high speed and in sterile conditions. Thereby facilitating the option to bring these cells back into culture, allowing creation of specific cell clones or performance of functional assays with the cells of interest. This sorter supports high-resolution analysis of up to 17 fluorescent markers simultaneously.

Union Biometrica Large Particle Biosorter:

Some objects are too large or too sensitive for conventional flow cytometry. Therefore, Union Biometrica has developed large particle flow cytometers for handling a wide range of object sizes (1 – 1500 µm). Examples of materials that can be analyzed and sorted with the Biosorter are plant protoplasts, seeds, organoids and whole organisms, including zebrafish embryos and C. elegans embryos and larvae.

Relative size and optical density are measured, as well as fluorescence intensities at three different wavelengths. Sorting and dispensing decisions are based on user-selected ranges using the FlowPilot™ Pro software. Of each object, an optical profile can be made, in which the location and intensity of all parameters can be graphically mapped. This feature makes it possible to distinguish between fluorescence in the head, middle, or tail of the organism.