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. 2021 Jun 18;9(6):e002894. doi: 10.1136/jitc-2021-002894

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© Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See http://creativecommons.org/licenses/by-nc/4.0/.

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Fine needle aspiration (FNA) enables single-cell RNA sequencing by multiplexing analysis using cell hashing. (A) FNA biopsies from four mice were labeled with DNA-barcoded antibody ‘hashtags’, pooled, sorted by FACS to enrich for immune cells and sequenced before demultiplexing and analysis. (B) UMAP embedded on hashtags colored by hashtag classification. (C) QC plot of hashtag classification versus counts of hashtag. Cells classified as HashtagA have higher counts for HashtagA, etc, while doublets present a mixed signal, and negative cells show few counts. (D) UMAP embedded on RNA colored by hashtag classification with putative cell types labeled. (E) Expression of the genes for the T cell marker CD3 and the B cell marker CD79a. UMAP, Uniform Manifold Approximation and Projection.