Bronchoalveolar Lavage Fluid Analysis

BALF was collected by lavaging lungs with 1.0 mL of sterile PBS. Cells were isolated by centrifugation, and total cell counts were determined using a Guava easyCyte HT cytometer (Millipore) and analyzed using Flowjo (version 10.4.2, Ashland, OR: Becton, Dickinson and Company, Ashland, OR, USA). Differential cell counts were obtained via cytospins using Hema3-stained (Fisher Scientific) total cells, on a minimum of 300 cells/animal. The differential cell counts were normalized to the total cell counts [3 (link)].

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Chamberlain N., Ruban M., Mark Z.F., Bruno S.R., Kumar A., Chandrasekaran R., Souza De Lima D., Antos D., Nakada E.M., Alcorn J.F, & Anathy V. (2022). Protein Disulfide Isomerase A3 Regulates Influenza Neuraminidase Activity and Influenza Burden in the Lung. International Journal of Molecular Sciences, 23(3), 1078.

Publication 2022

Guava easyCyte HT cytometer Hema3

Animal Cells Centrifugation Guava Sterile

Corresponding Organization : University of Vermont

Other organizations : Children's Hospital of Pittsburgh, University of Pittsburgh

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Variable analysis

independent variables

Lavaging lungs with 1.0 mL of sterile PBS

dependent variables

Total cell counts
Differential cell counts

control variables

Sterile PBS

controls

No positive or negative controls were explicitly mentioned.

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