Department of Chemistry, Columbia University, New York, NY, USA.
Nat Commun. 2021 Jun 7;12(1):3405. doi: 10.1038/s41467-021-23700-0.
Single-cell multiparameter measurement has been increasingly recognized as a key technology toward systematic understandings of complex molecular and cellular functions in biological systems. Despite extensive efforts in analytical techniques, it is still generally challenging for existing methods to decipher a large number of phenotypes in a single living cell. Herein we devise a multiplexed Raman probe panel with sharp and mutually resolvable Raman peaks to simultaneously quantify cell surface proteins, endocytosis activities, and metabolic dynamics of an individual live cell. When coupling it to whole-cell spontaneous Raman micro-spectroscopy, we demonstrate the utility of this technique in 14-plexed live-cell profiling and phenotyping under various drug perturbations. In particular, single-cell multiparameter measurement enables powerful clustering, correlation, and network analysis with biological insights. This profiling platform is compatible with live-cell cytometry, of low instrument complexity and capable of highly multiplexed measurement in a robust and straightforward manner, thereby contributing a valuable tool for both basic single-cell biology and translation applications such as high-content cell sorting and drug discovery.
单细胞多参数测量已逐渐被认为是系统理解生物系统中复杂分子和细胞功能的关键技术。尽管在分析技术方面进行了广泛的努力，但对于现有方法来说，仍然难以在单个活细胞中破译大量表型。在这里，我们设计了一个具有尖锐和相互可分辨的 Raman 峰的多路复用 Raman 探针面板，以同时定量单个活细胞的表面蛋白、内吞作用活性和代谢动力学。当将其与全细胞自发 Raman 微光谱法结合使用时，我们证明了该技术在各种药物干扰下对 14 重活细胞分析和表型分析的实用性。特别是，单细胞多参数测量能够进行强大的聚类、相关性和网络分析，并具有生物学见解。该分析平台与活细胞细胞术兼容，仪器复杂度低，能够以稳健和直接的方式进行高度多重测量，从而为基础单细胞生物学和转化应用（如高内涵细胞分选和药物发现）提供了有价值的工具。
