摘要
细胞异质性是生物系统复杂性的重要体现,在个体发育及疾病发生与发展过程中发挥着关键作用。然而,传统的细胞分析方法多基于大量细胞群体的平均信号,这种“平均化”处理方式会不可避免地掩盖稀有细胞亚群的特征和单细胞水平的细微差异,导致关键的异质性信息被忽视,难以真实反应单个细胞间的个体差异。因此,开展高通量的单细胞分析对于深入理解生命过程具有重要意义。近年来,随着生命科学研究向精细化和精准化方向不断推进,单细胞分析技术已成为科研热点。为了深入研究细胞异质性,建立一种基于电感耦合等离子体飞行时间质谱(ICP-TOF-MS)的单细胞多参数分析方法。在研究过程中,通过改造商用仪器的进样系统并优化关键分析参数,显著提升了单细胞分析的效率。结果表明,采用低流量同心雾化器和单通道雾室的高效进样系统(HSIS)可实现19.9%的单细胞进样效率,较传统同心雾化器与旋流雾室组成的标准系统(2.8%)提升了7倍。采用建立的方法对金属同位素标记的健康人全血经红细胞裂解后的样本进行多标记蛋白检测,验证了方法的可行性。建立的方法可实现高通量的单细胞蛋白组学分析,为深入解析免疫细胞异质性提供了有力工具,并有望应用于免疫细胞分化、肿瘤免疫微环境解析及免疫逃逸机制等方向的研究,为相关疾病的诊断、治疗和预后评估提供新的思路。
Cellular heterogeneity,a fundamental characteristic underlying biological complexity,is significantly influential in individual development,as well as in the initiation and progression of various diseases.However,traditional cell analysis techniques,which are based on population-averaged signals,are unable to precisely capture the subtle differences among individual cells.The crucial heterogeneity information is inevitably lost due to this averaging approach,thus impeding the accurate characterization of intercellular variations.As a result,highthroughput single-cell analysis has emerged as an indispensable tool for a more profound understanding of biological processes.In recent years,with the growing demand for precision and refinement in life sciences,single-cell analysis technologies have become a focal point of research interest.In this study,a novel multi-parametric singlecell analysis method was developed leveraging inductively coupled plasma time-of-flight mass spectrometry(ICPTOF-MS).Four distinct single-cell introduction systems were systematically compared and quantitatively evaluated to assess their performance.By modifying a commercial sample introduction system and optimizing key analytical parameters,a remarkable enhancement in single-cell analysis efficiency was achieved.Notably,an impressive single-cell introduction efficiency of 19.9%was demonstrated by the high-efficiency sample introduction system(HSIS),which features a low-flow concentric nebulizer integrated with a single-channel spray chamber.This figure represents a sevenfold improvement compared to the conventional system,which consists of a standard concentric nebulizer and a cyclonic spray chamber,with an efficiency of only 2.8%.Furthermore,two different sample delivery mechanisms,a syringe pump and a pressure-driven sample introduction device,were explored.Experimental results indicated that cell sedimentation issues were effectively mitigated and the risk of nebulizer clogging was substantially reduced by the pressure-driven system,which is equipped with a built-in selfmixing function.To validate the practicality of the proposed method,multiplexed protein detection was conducted on metal-isotope-labeled leukocytes derived from whole blood samples of healthy donors after red blood cell lysis.High-throughput single-cell proteomic analysis can be enabled by the developed method,which offers a powerful means to dissect immune cell heterogeneity.Great potential for advancing research in areas such as immune cell differentiation and tumor immune evasion is held by this breakthrough,thereby presenting new opportunities for the diagnosis,treatment,and prognosis of related diseases.
作者
田家乐
郑令娜
逄雨衡
鲁俊峰
王文敬
汪冰
王萌
丰伟悦
TIAN Jiale;ZHENG Lingna;PANG Yuheng;LU Junfeng;WANG Wenjing;WANG Bing;WANG Meng;FENG Weiyue(CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety,Institute of High Energy Physics,Chinese Academy of Sciences,Beijing 100049,China;University of Chinese Academy of Sciences,Beijing 100049,China;Beijing Youan Hospital,Capital Medical University,Beijing 100069,China)
出处
《中国无机分析化学》
北大核心
2026年第3期461-468,共8页
Chinese Journal of Inorganic Analytical Chemistry
基金
国家重点研发计划项目(2023YFF0612603)
北京市自然科学基金资助项目(Z230008,7232080)
国家自然科学基金资助项目(22474142)
首都临床特色诊疗技术研究及转化应用项目(Z221100007422063)
中国科学院高能物理研究所科技创新项目(E4545CU210)。
