美国哈佛大学Peng Yin和Sinem K. Saka等研究人员开发了Light-Seq，固定细胞和组织中生物分子的光定向原位条形码用于空间索引测序。这一研究成果2022年10月10日在线发表于《自然—方法学》上。

研究人员提出了Light-Seq策略，一种在固定细胞和组织中采用光定向DNA条形码对完整生物样本进行多重空间标引的方法。Light-Seq结合了空间靶向和快速光交联DNA条形码到互补DNA的原位一步DNA拼接反应，以创建汇集的、空间索引的测序库。这种光导向条形码能够在完整的固定组织样本中原位选择多个细胞群，根据位置、形态或蛋白质染色进行全转录组测序，不需要细胞分离。

将Light-Seq应用于小鼠视网膜切片，研究人员从三个细胞层中回收了数以千计的差异富集转录物，并从每个切片中仅4至8个单个细胞中发现了非常罕见的神经元亚型--多巴胺能无长突细胞的生物标志物。Light-Seq提供了一个可访问的工作流程，将原位成像、蛋白质染色与同一细胞的下一代测序相结合，使样本完整，以便测序后进一步分析。

附：英文原文

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

Author: Kishi, Jocelyn Y., Liu, Ninning, West, Emma R., Sheng, Kuanwei, Jordanides, Jack J., Serrata, Matthew, Cepko, Constance L., Saka, Sinem K., Yin, Peng

Issue&Volume: 2022-10-10

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

DOI: 10.1038/s41592-022-01604-1

Source: https://www.nature.com/articles/s41592-022-01604-1