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. 2021 Jun 2;8:674747. doi: 10.3389/fmolb.2021.674747

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Copyright © 2021 Hoyt.

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) and the copyright owner(s) 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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Assessment of TSA staining interference in detection of multiple markers within the same cellular compartment. Four membrane markers were chosen for this experiment which are known to have significant co-localization with one another: CD8 (Opal 520), CD3 (Opal 620), CD45RO (Opal 570), and CD45LcA (Opal 690). All reagents were double dispensed using the BOND RX and scanned using the Vectra Polaris. (A) Image with arrows indicates cells that display all four markers. (B) Additional representative images of cells displaying all four markers without any reduction in signal intensity demonstrating that TSA does not interfere with detection of three or more markers co-localized to the cell membrane.