The following examples illustrate unique applications which are available with flow cytometry instrumentation. Cell cycle analysis and immunophenotyping are the predominant applications performed in the facility, however assays measuring cellular function and viability have been conducted.
DNA analysis. Canine melanoma cells were fixed and permeabilized for subsequent Prodium Ioidide staining of DNA. Cell cycle analysis was performed and the results were compared to cell cycle analysis of canine peripheral blood mononuclear cells. The resultant DNA index was 1.1 illustrating tumor hyperploid. analyze flow cytometric data from remote computer labs within the university network. All computers in the facility are networked to Veterinary Medicine and LSU mainframes and data is readily transferred within the university.
NH4Cl lysed human peripheral blood. The scatterplot represents cell size (x-axis) versus internal granularity (y-axis) for peripheral blood leukocytes. A three part differential of lymphocytes, monocytes and granulocytes (predominantly neutrophils) are illustrated using a bivariate histogram. Lymphocytes are located in the polygon R1 and represent 32% of peripheral blood leukocytes. Red blood cells are lysed with NH4CL and platelets as well as ghost RBCs are eliminated based on size threshold.
Immunophenotyping. Within the lymphocyte population are various subsets as determined by anti-CD4-FITC and anti-CD8-PE staining. Dot display analysis illustrates the sample contains 48% CD4 T-lymphocytes (lower right quadrant) and 22% CD8 T-lymphocytes (upper left quadrant). The upper right quadrant contains dual stained CD4+CD8+ T-lymphocytes. The lower left quadrant contains dual negative cells which includes other subpopulations such as B-lymphocytes.