The post-thymic activation and development of T cells involves a heterogeneous population of cells at different stages of differentiation, including naïve, stem cell memory, central memory, effector memory and terminal effector T cells. These different T-cell subsets are themselves highly heterogeneous, and their identification and characterization require not one or a few but, rather, a […]
Scientists already know that MAIT ( Mucosal-associated invariant T) cells defend us against invading bacteria and fungi. Recently, a team of UK-based researchers discovered how MAIT cells also perform another of their vital functions: orchestrating the activity of other important cells in the immune system, such as dendritic cells (DCs). As you will see, Flow […]
In the study of rare cell types, Flow Cytometry is proposed to overcome the limitations of ELISA. By providing both phenotyping and functional data in the same assay, Flow Cytometry is well suited for investigating rare cell populations, especially cytokine producing cell subsets in an heterogeneous sample. Combined with the flexibility of Optibuild reagents for panel design, BD’s FACSCelesta™ allows you to perform advanced analysis of rare cells such as DCs and Tregs.
An important factor that must be considered for proper panel design is the sensitivity of the reagent. Sensitivity is influenced by the brightness of the fluorochrome and the data spread due to the presence of other antibody conjugates attached to the same cell. Not all fluorochromes have the same sensitivity due to differing brightness. Sensitivity is also impacted by the expression level of the markers being studied. Markers, or antigens, can be classified as primary, secondary or tertiary, depending on their levels of expression. Many candidate antibody-conjugate panels may need to be evaluated to determine which one optimally stains all the antigens under study. This is also important to determine which antibody conjugates reduce the sensitivity. Single stain controls to evaluate and correct for compensation must also be performed. Multicolor experiments can be frustrating but using the right panel can give wonderful results and it is important to invest time and effort into this.
Mahnke, Y.D, Roederer, M, Clin. Lab. Med; 27(3): 469, (2007)