Countess c10281

After 3–0 nylon ligations, 1 ml of physiological saline was instilled in the tracheal cannula (20 G) with subsequent aspiration with a syringe. This was repeated twice, yielding two samples from each animal according to previous methods [20 (link), 22 (link)] with some modification. The total number of cells was counted by an automated cell counter (Countess C10281; Invitrogen) under trypan blue stain. In addition, total leukocyte numbers and differential counts, lymphocytes, neutrophils, eosinophils, and monocytes, were detected by an automated hematology cell counter (Cell-DYN3700, Abbott Laboratories, Abbott Park, IL, USA).

BALF (approximately 0.7 mL) was collected by injecting saline (1 mL) through the tracheal cannula of the ligated right lobe followed by aspiration with a syringe. After staining the cells of BALF with trypan blue (Sigma-Aldrich; St. Louise, MO, USA), the total number of cells was counted using an automated cell counter (Model, Countess C10281; Invitrogen; Carlsbad, CA, USA). In addition, a Cell-DYN3700 hematology analyzer (Abbott Laboratories; Abbott Park, IL, USA) was used to count white blood cells and their specified cells (e.g., lymphocyte, neutrophil, eosinophil, and monocyte).