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. 2017 Oct 30;8:1398. doi: 10.3389/fimmu.2017.01398

Figure 4.

Figure 4

Nasal-associated lymphoid tissue (NALT) contamination results in the over estimation of leukocyte cells within the oral mucosa tissues. Oral mucosa was harvested from the maxilla of C57BL/6J mice, digested with collagenase D, and processed to obtain single-cell suspensions. Cells were stained with vitality dye Zombie NIR followed by rat anti-mouse CD45 mAb conjugated to PE to identify live leukocytes by flow cytometry. Representative dot plots generated by FlowJo analysis are shown. In three of the samples shown (A–C), marginal gingiva was harvested without disruption of the anterior hard palate as outlined in Figures 1 and 2. In a fourth sample (D), incision #2 was medial, landing over the NALT compartment in the anterior hard palate and exposing the oral mucosa to potential NALT contamination. The live leukocyte cell population (Zombie NIRlow, CD45+) is gated and the number of cells found in these gates is given. Comparable results were seen in two other similar experiments.