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Cell viability was determined by differential counting using the vital dye Janus Green; only suspensions with cell viability greater than 90% were used for inoculation.
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Neutrophil counts were by automated differential counts using the Sysmex XE-2100 (Sysmex Corporation, Kobe, Japan).
Whole blood was analyzed for WBC and differential counts using standard biochemical procedures.
Cell proliferation assays were performed by direct counting using a bacterial counting chamber (Hausser Scientific Partnership, PA, USA) and differential interference contrast microscopy.
The cells were also cytospun onto glass slides for differential counting by using Diff Quick stain (Dade Behring, Newark, DE, USA).
A portion of the pooled BALF was set aside for total cell count by hemacytometer and for differential cell count using a cytospin preparation with Wright stain.
Cell differentials were counted using smears prepared by Cytospin and stained with Diff-Quik (Sysmex, Kobe, Japan).
In further experiments, we observed a 47% increase in monocytes 5 h after a similar test workout for the NT group by using differential counting (p < 0.01; n = 6).
The cell numbers were counted using a hemocytometer, and differential cell counts were performed on slides prepared by cytocentrifugation at 250 rpm for 3 min and Diff-Quick staining.
Differential expression analysis was done on raw counts using the default settings in the DESeq package.
Cells were counted using a Neubauer chamber, and differential cell counts were performed in cytospin samples stained with Wright Giemsa.
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