Light-Seq: light-directed in situ barcoding of biomolecules in fixed cells and tissues for spatially indexed sequencing

• DOI: 10.1038/s41592-022-01604-1
• Publication Date: 2022-10-10T16:04:06Z
• Authors: Jocelyn Y. Kishi; Ninning Liu; Emma R. West; Kuanwei Sheng; Jack J. Jordanides; Matthew Serrata; Constance L. Cepko; Sinem K. Saka; Peng Yin
• Keywords: Illumina dye sequencing; DNA Barcoding

Abstract We present Light-Seq, an approach for multiplexed spatial indexing of intact biological samples using light-directed DNA barcoding in fixed cells and tissues followed by ex situ sequencing. Light-Seq combines spatially targeted, rapid photocrosslinking barcodes onto complementary DNAs with a one-step stitching reaction to create pooled, indexed sequencing libraries. This enables selection multiple cell populations tissue full-transcriptome based on location, morphology or protein stains, without cellular dissociation. Applying mouse retinal sections, we recovered thousands differentially enriched transcripts from three layers discovered biomarkers very rare neuronal subtype, dopaminergic amacrine cells, only four eight individual per section. provides accessible workflow combine imaging staining next generation the same leaving sample further analysis post-sequencing.