From simple to complex, CoImmune researchers connect the dots through the science of multi-color flow cytometry. Detection of cell surface molecules combined with intra cellular staining for function as defined by the expression of cytokines can reveal the immunological status pre and post treatment. A consistent and reproducible method for determining phenotype and function starts with optimized reagents and panels. You can choose an established panel design or a customized one to better suit your needs. Samples are acquired using a BD Bioscience LSRII Special Order System capable of detecting up to 18 different colors.
Objective gating strategies depend on correct identification of antibody-fluorochrome negative cells. Traditionally, this is achieved employing a ‘fluorescence minus one’ FMO strategy. During the development of the multi-color panel below, it was obvious that ‘unstained cells’ could not be used as a representative negative control. Fluorochrome-conjugated isotype control antibodies may be considered an alternative to an FMO. However, different antibody clones show variable activity concerning non-specific binding to cells. Therefore, a background set using an isotype control may not represent the non-specific binding of the test antibody, itself derived from a different antibody clone.