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Updated: Mar 3, 2026
Precision-cut Mouse Lung Slices to Visualize Live Pulmonary Dendritic Cells
Published on: April 5, 2017
Precision-cut Mouse Lung Slices to Visualize Live Pulmonary Dendritic Cells
Miranda R Lyons-Cohen1, Seddon Y Thomas1, Donald N Cook1
1Immunity, Inflammation, and Disease Laboratory, Division of Intramural Research, National Institute of Environmental Health Sciences, NIH.
Insights
Precision-cut Lung Slices (PCLS) allow visualization of immune cell locations and interactions within intact lung tissue. This method complements flow cytometry for studying lung inflammation and allergic diseases.
Area of Science:
- Immunology
- Cell Biology
- Respiratory Medicine
Background:
- Inhaled allergens and pathogens alter lung immune cell populations.
- Flow cytometry and chemotaxis assays offer limited insights into in vivo lung cell behavior.
- Understanding lung immune cell localization is crucial for studying respiratory diseases.
Purpose of the Study:
- To introduce and validate Precision-cut Lung Slices (PCLS) as a method for visualizing lung cell localization and interactions.
- To demonstrate the utility of PCLS in studying immune cell behavior within the complex lung environment.
- To highlight PCLS as a complementary technique to flow cytometry and qPCR for lung research.
Main Methods:
- Generating Precision-cut Lung Slices (PCLS) from live or fixed lung tissue.
- Staining PCLS with fluorescently tagged antibodies against various cell types.
- Visualizing stained PCLS using confocal microscopy.
Main Results:
- Successfully visualized diverse lung cell types including immune cells (dendritic cells, macrophages, neutrophils, T cells, B cells) and structural cells (lymphatic, endothelial, epithelial cells).
- Enabled visualization of cellular interactions, such as dendritic cell-T cell interactions, in a 3D lung context.
- Demonstrated PCLS can encompass large lung areas, up to an entire lobe cross-section.
Conclusions:
- PCLS provide a powerful tool for visualizing cell location and migration patterns within the intact lung.
- PCLS facilitate the study of cell-cell interactions in situ, offering insights into dynamic cellular events.
- Combined with other techniques, PCLS enhance the comprehensive understanding of lung inflammation and allergic diseases.
Abstract:
Inhalation of allergens and pathogens elicits multiple changes in a variety of immune cell types in the lung. Flow cytometry is a powerful technique for quantitative analysis of cell surface proteins on immune cells, but it provides no information on the localization and migration patterns of these cells within the lung. Similarly, chemotaxis assays can be performed to study the potential of cells to respond to chemotactic factors in vitro, but these assays do not reproduce the complex environment of the intact lung. In contrast to these aforementioned techniques, the location of individual cell types within the lung can be readily visualized by generating Precision-cut Lung Slices (PCLS), staining them with commercially available, fluorescently tagged antibodies, and visualizing the sections by confocal microscopy. PCLS can be used for both live and fixed lung tissue, and the slices can encompass areas as large as a cross section of an entire lobe. We have used this protocol to successfully visualize the location of a wide variety of cell types in the lung, including distinct types of dendritic cells, macrophages, neutrophils, T cells and B cells, as well as structural cells such as lymphatic, endothelial, and epithelial cells. The ability to visualize cellular interactions, such as those between dendritic cells and T cells, in live, three-dimensional lung tissue, can reveal how cells move within the lung and interact with one another at steady state and during inflammation. Thus, when used in combination with other procedures, such as flow cytometry and quantitative PCR, PCLS can contribute to a comprehensive understanding of cellular events that underlie allergic and inflammatory diseases of the lung.
