Sputum_Flow_Analysis

Analysis of the cellular content of sputum via flow cytometry in order to develop a diagnostic platform for lung cancer.
Conclusion:
Induced sputum samples were processed to collect single cell suspensions while removing debris through filtration. Sputum samples can be analyzed by flow cytometry while contaminating squamous epithelial cells (SECs) can be excluded from further analysis with the inclusion of a viability dye. Two macrophage populations were identified, and morphologies confirmed by sorting experiments, that can serve as verification that the sample was obtained from the lung as opposed to being saliva. We determined at least 0.75 million cells are needed for reliable analysis. The use of flow cytometry to analyze sputum-derived cells serves as a reliable, non-invasive means to monitor the health status of the lung.
Notes:
Three-day sputum collection from persons aged 55-74 with at least a 30 pack-year smoking history at high risk of developing lung cancer. LSR II cytometer was checked prior to each analysis session with BD FACSDiva™ CS&T Research beads. The LSR II PMTs are adjusted and checked prior to each use using SPHERO™ Rainbow Calibration Particles.