Flow Cytometric Assays to Quantify fHbp Expression and Detect Serotype Specific Capsular Polysaccharides on Neisseria...

Flow cytometry provides an automated analysis of bacteria passing in fluid suspension through a laser light beam. Bacteria are first treated with antibodies that bind to a specific target. These antibodies are tagged to fluorophores that fluoresce when passed through a laser beam. As the bacteria pass sequentially through the laser beam, they absorb and scatter the light in forward and side (90°) angles. The forward angle scatter is proportional to the size of the bacteria and the 90° angle side scatter is proportional to the internal structure (granularity). In addition, the tagged antibodies bound specifically to each bacteria, emit fluorescent light at defined wavelengths that can be collected and measured.Here we describe two flow cytometry based assays to measure expression levels of protein and polysaccharide on the surface of Neisseria meningitidis.