Nasal lavage samples were filtered, washed, centrifuged, resuspended, and counted in the same fashion as for oropharyngeal swab samples. Cells were stained in the same manner as described for PBMC, and equal numbers were analyzed by flow cytometry to determine the cell type and activation.
70 μm cell strainer
The 70 μm cell strainer is a laboratory tool used for filtration and separation purposes. It features a mesh with pore size of 70 micrometers, which allows for the effective removal of larger cell clumps or debris from cell suspensions while retaining the desired cell population.
Lab products found in correlation
4 protocols using 70 μm cell strainer
Oropharyngeal Swab and Nasal Lavage Processing
Nasal lavage samples were filtered, washed, centrifuged, resuspended, and counted in the same fashion as for oropharyngeal swab samples. Cells were stained in the same manner as described for PBMC, and equal numbers were analyzed by flow cytometry to determine the cell type and activation.
Comprehensive Organ Necropsies and CFU Quantification
Nasal and oropharyngeal sampling for cell analysis
Flow Cytometry Analysis of Immune Cells in Tissue Gels
surgically removed
from the animals after euthanasia. The gels were processed using mechanical
disruption to create a cell suspension that was then filtered through
a 70 μm cell strainer (Celltreat). To count total cells in the
gels, the single-cell suspensions were stained with anti-mouse CD45
and DAPI to count live leukocytes. The cells were mixed with CountBright
Absolute Counting beads (ThermoFisher) and analyzed on a LSRFortessa
X-20 flow cytometer (BD Biosciences). To analyze neutrophil and other
myeloid cell counts in the gels, the single-cell suspensions were
incubated with anti-CD16/CD32 (produced in house from 2.4G2 hybridoma)
to block Fc receptor binding and then stained with anti-mouse CD45,
CD3, CD19, CD11c, Ly6G, CD11b, and MHCII in FACS buffer (2 mM EDTA,
2% FBS in PBS) for 30 min on ice. Dead cells were excluded by DAPI
staining. Cells were acquired on a LSRFortessa X-20, and fcs files
were analyzed using FlowJo 10 (FlowJo LLC). See
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