Improving the Quality and Reproducibility of Flow Cytometry in the Lung. An Official American Thoracic Society Workshop Report

Robert M Tighe, Elizabeth F Redente, Yen-Rei Yu, Susanne Herold, Anne I Sperling, Jeffrey L Curtis, Ryan Duggan, Suchitra Swaminathan, Hideki Nakano, William J Zacharias, William J Janssen, Christine M Freeman, Ryan R Brinkman, Benjamin D Singer, Claudia V Jakubzick, Alexander V Misharin, Robert M Tighe, Elizabeth F Redente, Yen-Rei Yu, Susanne Herold, Anne I Sperling, Jeffrey L Curtis, Ryan Duggan, Suchitra Swaminathan, Hideki Nakano, William J Zacharias, William J Janssen, Christine M Freeman, Ryan R Brinkman, Benjamin D Singer, Claudia V Jakubzick, Alexander V Misharin

Abstract

Defining responses of the structural and immune cells in biologic systems is critically important to understanding disease states and responses to injury. This requires accurate and sensitive methods to define cell types in organ systems. The principal method to delineate the cell populations involved in these processes is flow cytometry. Although researchers increasingly use flow cytometry, technical challenges can affect its accuracy and reproducibility, thus significantly limiting scientific advancements. This challenge is particularly critical to lung immunology, as the lung is readily accessible and therefore used in preclinical and clinical studies to define potential therapeutics. Given the importance of flow cytometry in pulmonary research, the American Thoracic Society convened a working group to highlight issues and technical challenges to the performance of high-quality pulmonary flow cytometry, with a goal of improving its quality and reproducibility.

Keywords: cells; flow cytometry; lung biology; reproducibility.

Figures

Figure 1.
Figure 1.
Effect of apoptotic/necrotic cells and cell debris on flow cytometry plots with and without use of a viability dye. The red circle highlights a population of nonviable cells that are identified when a viability dye is not used in the staining conditions. Representative sample of a murine lung tissue digestion. FSC = forward scatter; SSC = side scatter.

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