Multiparameter FACS analysis.

The purpose of this unit is three-fold: (1) toprovide a brief historical perspective to illustrate the demands for this technology and to lay the foundation for its application; (2) to explain the hurdles that were surmounted to achieve the current level of multiparametric analysis which serves to alert researchers to potential problems they may encounter when they either bring this technology to their own laboratories, or when they use extant instrumentation in another laboratory; and (3) to illustrate some of the complexities that arise even when all of the major technical hurdles have been overcome. The majority of the unit is focused on "high-end" multiparameter analysis (termed "polychromatic flow cytometry," or PFC); i.e., experiments utilizing more than four fluorescence detectors (in addition to scattered light parameters).