Flow cytometry panel design is a critical step in developing a flow cytometry protocol, but this can be a time consuming process, especially if you are working with a panel using four colors or more. Commercially available fluorescently-labeled antibodies are often species-specific, but some antibodies recognize highly conserved target epitopes and can be used to label cells from different species. Incorporating these cross-reactive antibodies into a flow cytometry panel can save you time as you work up panels for different species, be they mice, non-human primates (NHP), or humans. Consider these factors when looking for or using a cross-reactive antibody.
Flow Cytometry Blog
Flow cytometry is a powerful tool because it allows users to analyze the characteristics of millions of cells with relative speed and precision. A single cell suspension of fluorescently labeled sample travels through the cytometer for excitation by lasers, and the emitted light photons are measured by different detectors. Having a single cell suspension is essential to measuring cell fluorescence accurately, and many types of cell or tissue samples must be specially processed to make this suspension. Two different methods can be used to process samples into single cell suspensions--mechanical or enzymatic dissociation. This processing step is typically carried out before cells are stained and both methods have benefits and caveats.
The emergence of biologic-based therapeutics has transformed the treatment of cancer, autoimmune disease and other disorders. These biologically active molecules can alter the immune response and change disease trajectories, so many of the tests that are used to assess their function must examine biological functions. These molecules typically bind to specific receptors on immune cells, which may block or activate a specific response depending on the mode of action.
Drugs and biologics in the research pipeline must undergo stringent preclinical toxicology and safety assessments before use in a clinical trial. Most traditional preclinical toxicology screenings include testing in animal models in order to define toxicological and pharmacological parameters that are critical to determining appropriate dosing such as maximum tolerated dose (MTD).
T cells and NK cells deploy potent immune defenses through the release of lysosomal granule contents such as granzymes and perforin. Lysosomes containing cytotoxic granules move to the cell surface and fuse with the plasma membrane to release their contents upon specific receptor engagement between a target cell and cytotoxic cell. This lysosomal fusion results in lysosomal-associated membrane glycoproteins (LAMPs), including CD107a (LAMP-1) and CD107b (LAMP-2), to be presented on the cell surface.