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Cell “hashing” with barcoded antibodies enables multiplexing and doublet detection for single cell genomics

View ORCID ProfileMarlon Stoeckius, View ORCID ProfileShiwei Zheng, View ORCID ProfileBrian Houck-Loomis, View ORCID ProfileStephanie Hao, View ORCID ProfileBertrand Z. Yeung, View ORCID ProfilePeter Smibert, View ORCID ProfileRahul Satija
doi: https://doi.org/10.1101/237693
Marlon Stoeckius
1Technology Innovation Lab, New York Genome Center, New York, NY
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Shiwei Zheng
2New York Genome Center, New York, NY; NYU Center for Genomics and Systems Biology, New York, NY
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Brian Houck-Loomis
1Technology Innovation Lab, New York Genome Center, New York, NY
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Stephanie Hao
1Technology Innovation Lab, New York Genome Center, New York, NY
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Bertrand Z. Yeung
3BioLegend Inc., San Diego, CA
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Peter Smibert
1Technology Innovation Lab, New York Genome Center, New York, NY
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  • For correspondence: psmibert@nygenome.org rsatija@nygenome.org
Rahul Satija
2New York Genome Center, New York, NY; NYU Center for Genomics and Systems Biology, New York, NY
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  • ORCID record for Rahul Satija
  • For correspondence: psmibert@nygenome.org rsatija@nygenome.org
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ABSTRACT

Despite rapid developments in single cell sequencing technology, sample-specific batch effects, detection of cell doublets, and the cost of generating massive datasets remain outstanding challenges. Here, we introduce cell “hashing”, where oligo-tagged antibodies against ubiquitously expressed surface proteins are used to uniquely label cells from distinct samples, which can be subsequently pooled. By sequencing these tags alongside the cellular transcriptome, we can assign each cell to its sample of origin, and robustly identify doublets originating from multiple samples. We demonstrate our approach by pooling eight human PBMC samples on a single run of the 10x Chromium system, substantially reducing our per-cell costs for library generation. Cell “hashing” is inspired by, and complementary to, elegant multiplexing strategies based on genetic variation, which we also leverage to validate our results. We therefore envision that our approach will help to generalize the benefits of single cell multiplexing to diverse samples and experimental designs.

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Posted December 21, 2017.
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Cell “hashing” with barcoded antibodies enables multiplexing and doublet detection for single cell genomics
Marlon Stoeckius, Shiwei Zheng, Brian Houck-Loomis, Stephanie Hao, Bertrand Z. Yeung, Peter Smibert, Rahul Satija
bioRxiv 237693; doi: https://doi.org/10.1101/237693
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Cell “hashing” with barcoded antibodies enables multiplexing and doublet detection for single cell genomics
Marlon Stoeckius, Shiwei Zheng, Brian Houck-Loomis, Stephanie Hao, Bertrand Z. Yeung, Peter Smibert, Rahul Satija
bioRxiv 237693; doi: https://doi.org/10.1101/237693

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