20241120 HannanEtAl Mirror for FigShare.zip

Sample multiplexing in flow cytometry is a powerful technique which allows for reduction of error, inclusion of control samples for batch effect correction, and reduction in both time and consumable usage. Current industry standard for barcoding in mass cytometry is an intracellular reagent, which requires fixation and permeabilization of sample prior to barcoding. We developed a barcode using the ubiquitous and well-tolerated membrane labeling lectin, wheat germ agglutinin. This barcode effectively labels all tested cell types, both live and fixed. We determine that barcode yields, or the ratio of debarcoded cells to total input cells, is stable in live pooled sample for at least an hour. This barcode does not show differential performance across major PBMC lineages. Thus, this universal wheat germ agglutinin-based barcode represents an advance in gentle, non-reactive cell surface barcoding for live cells.

流式细胞术中的样本复用是一项强大技术，可降低误差、纳入对照样本以校正批次效应，并减少时间与耗材使用量。当前质谱流式细胞术中的行业标准条形码技术采用细胞内试剂，需在条形码标记前对样本进行固定与透化处理。我们利用普遍存在且耐受性良好的膜标记凝集素——小麦胚芽凝集素（wheat germ agglutinin）开发了一种条形码。该条形码可有效标记所有测试的细胞类型，包括活细胞与固定细胞。我们发现，条形码产率（即解条形码细胞与总输入细胞的比值）在混合活样本中至少一小时内保持稳定。该条形码在外周血单个核细胞（PBMC）主要谱系间未表现出差异性能。因此，这种基于小麦胚芽凝集素的通用条形码技术，代表了活细胞温和、非反应性细胞表面条形码技术的一项进步。