Department of Pathology, School of Medicine, Stanford University, Palo Alto, CA, USA.
Department of Neurology, UCSF, San Francisco, CA, USA.
Sci Rep. 2018 Jul 17;8(1):10770. doi: 10.1038/s41598-018-28791-2.
Single-cell barcoding enables the combined processing and acquisition of multiple individual samples as one. This maximizes assay efficiency and eliminates technical variability in both sample preparation and analysis. Remaining challenges are the barcoding of live, unprocessed cells to increase downstream assay performance combined with the flexibility of the approach towards a broad range of cell types. To that end, we developed a novel antibody-based platform that allows the robust barcoding of live human cells for mass cytometry (CyTOF). By targeting both the MHC class I complex (beta-2-microglobulin) and a broadly expressed sodium-potassium ATPase-subunit (CD298) with platinum-conjugated antibodies, human immune cells, stem cells as well as tumor cells could be multiplexed in the same single-cell assay. In addition, we present a novel palladium-based covalent viability reagent compatible with this barcoding strategy. Altogether, this platform enables mass cytometry-based, live-cell barcoding across a multitude of human sample types and provides a scheme for multiplexed barcoding of human single-cell assays in general.
单细胞标记技术能够将多个单独的样本合并处理和采集。这最大限度地提高了检测效率，并消除了样本制备和分析过程中的技术变异性。仍然存在的挑战是对活的、未经处理的细胞进行标记，以提高下游检测性能，同时保持该方法对广泛的细胞类型的灵活性。为此，我们开发了一种新的基于抗体的平台，允许对用于质谱流式细胞术（CyTOF）的活的人类细胞进行强大的标记。通过用铂缀合抗体靶向 MHC Ⅰ类复合物（β2-微球蛋白）和广泛表达的钠钾 ATP 酶亚基（CD298），可以在同一个单细胞检测中将人类免疫细胞、干细胞和肿瘤细胞进行多重标记。此外，我们还介绍了一种与该标记策略兼容的新型基于钯的共价细胞活力试剂。总的来说，该平台能够在多种人类样本类型中进行基于质谱流式细胞术的活细胞标记，并为一般的人类单细胞检测的多重标记提供了一种方案。
