Quality-Controlled Sputum Analysis by Flow Cytometry, Journal of Visualized Experiments (JoVE), August 9, 2021

Marcia Grayson, Shao-Chiang Lai, Lydia H. Bederka, Patricia Araujo, Jamila Sanchez, Xavier T. Reveles, Vivienne I. Rebel, Jennifer Rebeles

The paper presents an efficient and effective method to analyze sputum on a flow cytometric platform, allowing for the development of high-throughput diagnostic tests for lung cancer and other lung diseases. [Jump] A gating strategy that describes the exclusion of debris, dead cells (including SECs) and cell doublets is presented here. Further, this work also explains how to analyze viable, single sputum cells based on a cluster of differentiation (CD)45 positive and negative populations to characterize hematopoietic and epithelial lineage subsets. A quality control measure is also provided by identifying lung-specific macrophages as evidence that a sample is derived from the lung and is not saliva. Finally, it has been demonstrated that this method can be applied to different cytometric platforms by providing sputum profiles from the same patient analyzed on three flow cytometers; Navios EX, LSR II, and Lyric. Furthermore, this protocol can be modified to include additional cellular markers of interest. A method to analyze an entire sputum sample on a flow cytometric platform is presented here that makes sputum amenable for developing high-throughput diagnostics of lung disease.

Quality-Controlled Sputum Analysis by Flow Cytometry | Protocol (jove.com) 

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