The flow cytometry laboratory consists of two FACScans and a FACStar Plus, all of which are on a thin wire ethernet using TCP/IP. There are three other FACScan flow cytometers within CBER; they all have some level of network capability. Together these laboratories make up the CBER/FDA flow cytometry consortium. The Section of Flow & Image Cytometry is concerned with desk-top publishing of flow cytometric data, development of necessary software programs to carry this out in the Macintosh graphics user interface environment, and quantitative flow cytometry as it relates to our regulatory needs. The next area of data analysis in flow cytometry will consist of a comparison of cluster analysis programs. The major program concerns the etiology and pathogenesis of common and familial B cell lymphocytic leukemia (B-CLL). This has consisted of an evaluation of two and three color surface immunofluorescence, cell cycle analysis and sorting of B cells for PCR analysis of monoclonality. This latter activity is being undertaken as part of a linkage analysis study in familial B-CLL. It is designed to detect early preclinical CLL in third and fourth generation family members to facilitate the formal genetic linkage analysis. Presently, we use E-rosette depletion and monocyte depletion to prepare enriched B lymphocyte preparations. Oligomeric primers predicted from conserved sequences in the variable (VH) and joining (JH) regions are used in conjunction with another set of oligomeric primers in V-region families (VH1 through VH6) that are used on clonal evaluation. A mouse colony has been established to evaluate a murine model of B-CLL. Image analysis is being carried out to examine the morphological heterogeneity of lymphocytes seen on conventionally prepared blood films as a function of time. We plan to add optical sectioning of leukemic lymphocytes, and develop multi-media patient records. Our study with familial CLL is coordinated with the Family Studies Section outpatient clinic on the third floor of the Clinical Center. Bone marrow and lymph node biopsies, blood films, and immunoperoxidase assays are reviewed in the Hematopathology Section of the NCI.
