Chen Kangfu, Wang Zongjie
Department of Biomedical Engineering, McCormick School of Engineering, Northwestern University, Evanston, IL 60208, USA.
Chan Zuckerberg Biohub Chicago, Chicago, IL 60607, USA.
Methods Protoc. 2023 Sep 4;6(5):80. doi: 10.3390/mps6050080.
Advancements in single-cell-related technologies have opened new possibilities for analyzing rare cells, such as circulating tumor cells (CTCs) and rare immune cells. Among these techniques, single-cell proteomics, particularly single-cell mass spectrometric analysis (scMS), has gained significant attention due to its ability to directly measure transcripts without the need for specific reagents. However, the success of single-cell proteomics relies heavily on efficient sample preparation, as protein loss in low-concentration samples can profoundly impact the analysis. To address this challenge, an effective handling system for rare cells is essential for single-cell proteomic analysis. Herein, we propose a microfluidics-based method that offers highly efficient isolation, detection, and collection of rare cells (e.g., CTCs). The detailed fabrication process of the micropillar array-based microfluidic device is presented, along with its application for CTC isolation, identification, and collection for subsequent proteomic analysis.
单细胞相关技术的进步为分析罕见细胞(如循环肿瘤细胞(CTC)和罕见免疫细胞)开辟了新的可能性。在这些技术中,单细胞蛋白质组学,特别是单细胞质谱分析(scMS),因其无需特定试剂即可直接测量转录本的能力而备受关注。然而,单细胞蛋白质组学的成功在很大程度上依赖于高效的样品制备,因为低浓度样品中的蛋白质损失会对分析产生深远影响。为应对这一挑战,一种有效的罕见细胞处理系统对于单细胞蛋白质组学分析至关重要。在此,我们提出一种基于微流控的方法,该方法可实现对罕见细胞(如CTC)的高效分离、检测和收集。本文介绍了基于微柱阵列的微流控装置的详细制造过程,以及其在CTC分离、鉴定和收集以用于后续蛋白质组学分析中的应用。