Sputum_analysis_LSRII_cytometer

To determine whether our method for sputum processing can be applied to different flow cytometric platforms. Conclusion: Using our methods, we were able to successfully dissociate sputum into a single cell suspension into live, viable cells that were stained with blood and epithelial lineage markers and analyzed by three flow cytometric platforms. 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. This experiment was used as a proof of concept of our protocol for a methods manuscript. No treatments or experimental conditions were applied to the samples. 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. The presence of alveolar macrophages was verified to determine the sample derives from the lung.