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

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Copyright © 2017 Bittner-Eddy, Fischer, Tu, Allman and Costalonga.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Processed oral mucosa samples are contaminated with on average 25% circulating leukocytes. Anesthetized C57BL/6J mice were given anti-CD45:FITC mAb as described in Figure 7 and euthanized at 3 min. Single-cell suspensions were processed from oral mucosa samples from single mice, stained with vitality dye Zombie NIR followed by rat anti-mouse CD45:PE to identify live immune cells by flow cytometry. (A) Gating strategy used to identify live interstitial (Zombie NIR^low, CD45:PE⁺, CD45:FITC⁻) and circulating (Zombie NIR^low, CD45:PE⁻, CD45:FITC⁺) immune cell populations. Percentage of cells within a drawn gate is given. (B) Distribution of immune cells from oral mucosa samples into interstitial or circulating leukocytes fractions. Data are from three independent experiments (n = 20).