Flow Cytometry Core Applications

Immunophenotyping: It is one of the most common uses of flow cytometry, uses fluorescence conjugated antibodies to detect specific antigens expressed both on cell surface or internally.

Cell cycle and ploidy analysis: DNA probes (like PI, propidium iodide) intercalates into cellular DNA and the fluorescent intensity indicates the number of chromosomes in each cell. Normal cells have diploid DNA content (cell cycle picture).

Apoptosis analysis: Apoptosis can be detected on flow cytometer by different functional indicators like:

• Fluorescence-conjugated Annexin-V and PI/7-AAD two-color staining
• DNA analysis to look at subG₀ peak
• TUNEL assay to detect DNA fragments
• Hoechst staining to look for DNA conformation change

Cell proliferation: Detect generations of cells by using CFSE (Carboxyfluorescein succinimidyl ester) dye which incorporates into the cell membrane and passed from one generation to the next. The amount of dye in proliferating cells is halved with each successive division.

Fluorescent proteins: These are used as a maker/reporter to determine transfection efficiency or gene expression on FLOW (e.g., GFP ).

Multiplex Assay: Using the antibody-coupled Cytometric bead array (CBA) Capture beads to measure multiple analytes (proteins) in a single sample simultaneously and quantitatively. See 12-flex CBA assay data on our Aria.

Cellular function Probes: Include fluorescent probes to detect intracelluar ions (Ca2+, Mg2+ and K+), intracelluar pH, and membrane potential

Cell Sorting: The interested cell population can be separated from a mixture of cells according to specific property (labeling) and collected for further analysis (See the pre-sort and post-sort data ).

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