Mouse Lung Lavage and Cell Analysis

The lung of each mouse was lavaged with 3 × 0.5 ml of sterile saline according to previously described^{59 (link),60 (link)}. The BALF was centrifuged at 250 × g for 10 min. The pellet was resuspended with 1 ml of sterile saline, and then the cells were mounted on a slide by cytospin centrifuge (Hettich ROTOFIX 32 A) at 1000 rpm for 3 min. Slides were air-dried and stained with Hema-3 Stain Kit (Fisher Scientific Company, Kalamazoo, MI) for analysis with Nikon Eclipse TE2000-U research microscope (Nikon, Melville, NY).

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Du J., Abdel-Razek O., Shi Q., Hu F., Ding G., Cooney R.N, & Wang G. (2018). Surfactant protein D attenuates acute lung and kidney injuries in pneumonia-induced sepsis through modulating apoptosis, inflammation and NF-κB signaling. Scientific Reports, 8, 15393.

Publication 2018

Hema-3 Stain Kit Nikon Eclipse TE2000-U research microscope

Cells Hema Lung Microscope Mouse Saline Stain Sterile

Corresponding Organization :

Other organizations : SUNY Upstate Medical University, Renmin Hospital of Wuhan University, Wuhan University

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Variable analysis

independent variables

The lungs of each mouse were lavaged with 3 × 0.5 ml of sterile saline

dependent variables

The bronchoalveolar lavage fluid (BALF) was centrifuged at 250 × g for 10 min
The pellet was resuspended with 1 ml of sterile saline
The cells were mounted on a slide by cytospin centrifuge (Hettich ROTOFIX 32 A) at 1000 rpm for 3 min
Slides were air-dried and stained with Hema-3 Stain Kit (Fisher Scientific Company, Kalamazoo, MI) for analysis with Nikon Eclipse TE2000-U research microscope (Nikon, Melville, NY)

control variables

The lungs were lavaged with sterile saline
The centrifugation and resuspension steps were performed under standardized conditions
The cytospin centrifugation and slide preparation were carried out under controlled settings
The staining and microscopic analysis were performed using standardized techniques and equipment

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