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ZipSeq : Barcoding for Real-time Mapping of Single Cell Transcriptomes

By Kenneth H. Hu, John P Eichorst, Chris S. McGinnis, David M Patterson, Eric D. Chow, Kelly Kersten, Stephen C Jameson, Zev J. Gartner, Arjun A Rao, Matthew F Krummel

Posted 04 Feb 2020
bioRxiv DOI: 10.1101/2020.02.04.932988 (published DOI: 10.1038/s41592-020-0880-2)

Spatial transcriptomics seeks to integrate single-cell transcriptomic data within the 3-dimensional space of multicellular biology. Current methods use glass substrates pre-seeded with matrices of barcodes or fluorescence hybridization of a limited number of probes. We developed an alternative approach, called ZipSeq, that uses patterned illumination and photocaged oligonucleotides to serially print barcodes (Zipcodes) onto live cells within intact tissues, in real-time and with on-the-fly selection of patterns. Using ZipSeq, we mapped gene expression in three settings: in-vitro wound healing, live lymph node sections and in a live tumor microenvironment (TME). In all cases, we discovered new gene expression patterns associated with histological structures. In the TME, this demonstrated a trajectory of myeloid and T cell differentiation, from periphery inward. A variation of ZipSeq efficiently scales to the level of single cells, providing a pathway for complete mapping of live tissues, subsequent to real-time imaging or perturbation.

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