Objective quantitative analysis of eosinophils and bronchial epithelial cells - 18 Downloads
Objective quantitative analysis of eosinophils and
bronchial epithelial cells in induced sputum by laser scanning cytometry
G Woltmann, RJ Ward, FA Symon, DA Rew, I D Pavord, A J Wardlaw
Abstract
Background-Sputum induction is an important non-invasive technique for measuring airway inflammation in asthma. Cell numbers are often too low for flow cytometric analysis. Laser scanning cytometry (LSC) is a novel technique that allows objective multicolour fluorescence analysis of cells on a microscope slide.
Methods
LSC was used to determine sputum eosinophil and bronchial epithelial cell counts. We first confirmed that we could measure eosinophil counts accurately in peripheral blood using a-major basic protein (MBP) immunofluorescent staining. Sputum induction was performed according to standard protocols. Sputum samples from eight normal controls and 12 asthmatic patients were analysed by LSC and manual counting by two independent observers. Octospot cytospins were fixed and stained with mousea-human-MBP monoclonal antibody or mouse-a-human-cytokeratin antibody and goat-a-mouse Oregon Green conjugated second antibody.
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