A semi-automated immunofluorescent microscopy system for enumeration of immunoglobulin producing cells

A computer-based video microscopy (semi-automated) system (SAS) has been developed for use in characterizing immune responses in mice. The system employs a fluorescent microscope, intensified CCD camera system, autoscanning stage, PC and other PC-based hardware and software for image capture, analysis and processing. In addition, customized algorithms were developed for the sorting and selection of true image information from artifacts, and quantitation. A specially developed immunofluorescent assay is used to label or tag antibodies (immunoglobulins) secreted by murine immune (B) cells with a fluorescent marker. The enumeration (counting) of the number of these fluorescent immune cell footprints or spot forming cells (SFCs) is used by immunologists to evaluate the effectiveness of immune responses and vaccines in providing immunity. Current studies involve comparing the conventional method of SFC eye counts from experienced research personnel with counts generated by the SAS. Results from preliminary studies have indicated that the SAS can perform at least as well as trained research personnel in enumerating the number of immunoglobulin SFCs. Full implementation of such an automated system may be invaluable to researchers in immunology in terms of the time and tedium saved for sample analysis and SFC enumeration