Cellular and Biochemical Analysis of Bronchoalveolar Lavage Fluid from Murine Lungs

Methods Mol Biol. 2021:2223:201-215. doi: 10.1007/978-1-0716-1001-5_15.

Abstract

Bronchoalveolar lavage (BAL) is a technique used to collect the contents of the airways. The fluid recovered, called BAL fluid (BALF), serves as a dynamic tool to identify various disease pathologies ranging from asthma to infectious diseases to cancer in the lungs. A wide array of tests can be performed with BALF, including total and differential leukocyte counts (DLC), enzyme-linked immunosorbent assays (ELISA) or flow-cytometric quantitation of inflammatory mediators, such as cytokines, chemokines and adhesion molecules, and assessment of nitrate and nitrite content for estimation of nitric oxide synthase (NOS) activity. Here, we describe a detailed procedure for the collection of BALF for a variety of downstream usages, including DLC by cytological and flow-cytometry-based methods, multiplex cytokine analysis by flow cytometry, and NOS activity analysis by determining nitrate and nitrite levels.

Keywords: Cytokines; Differential leukocyte count; Flow cytometry; Nitric oxide synthase; Tracheostomy.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Basophils / cytology
  • Basophils / metabolism
  • Bronchoalveolar Lavage Fluid / chemistry
  • Bronchoalveolar Lavage Fluid / cytology*
  • Bronchoalveolar Lavage Fluid / immunology
  • Cytokines / immunology
  • Cytokines / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Eosinophils / cytology
  • Eosinophils / metabolism
  • Flow Cytometry / methods*
  • Humans
  • Leukocyte Count
  • Lung / cytology*
  • Lung / immunology
  • Lung / metabolism
  • Macrophages, Alveolar / cytology*
  • Macrophages, Alveolar / metabolism
  • Mice
  • Monocytes / cytology
  • Monocytes / metabolism
  • Neutrophils / cytology*
  • Neutrophils / metabolism
  • Nitric Oxide Synthase / immunology
  • Nitric Oxide Synthase / metabolism
  • Tracheostomy / methods

Substances

  • Cytokines
  • Nitric Oxide Synthase