Flow Cytometric Methods for Assaying Damage to Respiratory Tract Cells

摘要

This paper summarizes results of experiments designed to develop automated flow-analysis assay methods for discerning damage to exfoliated respiratory tract cells in model test animals exposed by inhalation to physical and chemical agents associated with the production of synthetic fuels from oil shale, the specific goal being the determination of atypical changes in exposed lung macrophages and epithelial cells. Animals were exposed to oil shale particulates (raw and spent), silica, and ozone, and respiratory tract cells were obtained by lavaging the lungs with normal saline. Cell samples were stained with fluorescent dyes specific for different biochemical parameters and analyzed as they flowed through a chamber intersecting a laser beam(s) of exciting light where sensors measured fluorescence and light scatter (cell size) on a cell-by-cell basis. Cellular parameters proportional to optical signals were displayed as frequency distribution histograms. Cells also were separated according to cytological features and identified. The basic features of the methodology are presented, along with examples of results that illustrate characterization and analysis of normal and exposed respiratory tract cells based on DNA content, total protein, size, and phagocytic activity. (ERA citation 04:052233)