Flow Analysis and Separation of Exfoliated Respiratory Tract Cells

摘要

This paper describes results of preliminary experiments designed to develop automated flow-analysis methods for determining damage to respiratory tract cells in humans exposed by inhalation of physical and chemical toxic agents, the specific goal being the determination of early atypical changes in exposed lung epithelium using the Syrian hamster as a model test system. Hamster respiratory cell samples composed of macrophages, leukocytes, ciliated columnar, and basal undifferentiated cells were obtained by lavaging the lungs with physiological saline. Cell samples stained with fluorescent dyes specific for various biochemical parameters were analyzed in liquid suspension as they flowed through a chamber intersecting a laser beam of exciting light. Sensors measured the fluorescence and light-scatter optical signals on a cell-by-cell basis. Cellular parameters proportional to optical signals (e.g., enzymatic esterase activity, cell size, DNA content, protein, nuclear and cytoplasmic diameter) were displayed as frequency distribution histograms. Cells were also separated electronically and identified microscopically. The basic operating features of the technology are presented, along with representative examples of results which illustrate initial characterization of normal exfoliated respiratory tract cells. (ERA citation 02:053062)