Controls are the most important part of any multicolor flow cytometry experiment. When using multiple colors it is necessary to have the right controls to compensate for emission spectral overlap.  Fluorescence minus one (FMO) controls – cells or beads stained with all but one of the fluorophore – are used to determine the spillover for each spectral emission.  A mathematical method is then applied to compensate for the overlap from each fluorophore.

Each new color in a panel increases the chance of non-specific spillover into almost all other channels.  FMO tubes negative for one the fluorophores in the panel and gating is used to measure fluorescence from the target antibody.   FMO staining critical for removing the discrepancies that remain after compensating for all the colors. FMO is used in addition to compensation controls to identify gating boundaries and differentiate spillover emission when fluorophores are excited by multiple lasers.

Flow cytometry can be used to gather a plethora of experimental data. Power and versatility have made flow cytometry a cornerstone in modern vaccinology and immunology: an essential tool for evaluating drug and vaccine efficacy. It is also a complex tool with many experimental design challenges to generating reproducible and publishable data.

IBT Bioservice can design and execute your studies with a variety of panels using up to twenty-three colors. We have panels validated using mouse, human and non-human primate samples. We offer immunophenotyping of T-lymphocyte and B-lymphocytes, detailed multicolor evaluation of T-lymphocyte response to antigens and much more. Our expert staff is always available to guide you on markers and fluorophores selection for custom analysis of the specific cell subset of interest for your project. To save money and time visit our website or emails your questions to Services@IBTBioservices.com