单细胞分析用流式质谱仪的研制与表征

Development and Characterization of a Mass Cytometer for Single-Cell Analysis

  • 摘要: 流式细胞术存在检测通道受限(通常<12个)和荧光光谱重叠等技术瓶颈,而基于质谱检测原理的流式质谱技术通过引入金属同位素标记的方法,实现了检测通道的显著扩展(>40个)和检测通道间信号干扰的有效抑制。然而,进口设备的高昂成本严重制约了该技术向科学研究及临床检验等领域的推广应用。本研究成功研制了一套国产化的流式质谱仪,由气动进样模块、电感耦合等离子体(ICP)离子源、四极杆离子传输系统、飞行时间质量分析器(TOF MS)和高频数据采集系统等核心部件组成,采用多元素金属标准溶液对仪器性能进行系统表征。结果表明,自制仪器的全质量范围分辨率超过550(FWHM);在30 μL/min进样流速下,灵敏度达到9×105 cps/pg(159Tb+);对Cs、Tb、Ir的检出限分别为0.4、0.2、0.04 ng/L;8 h质量稳定性测试显示,质量偏移量<0.1 u;细胞表面蛋白表达的检测结果与国际主流仪器(Helios)具有高度一致性。本研究成果填补了国内高端质谱流式细胞分析仪器的技术空白,为推进单细胞多参数研究提供了重要的技术支撑。

     

    Abstract: Flow cytometry is widely used for single-cell analysis but is constrained by a limited number of detection channels (typically<12) and by spectral overlap between fluorophores. Mass cytometry, based on mass spectrometric detection principles, addresses these limitations by employing metal isotope labeling, enabling a substantial expansion of detection channels (>40) and effectively reducing inter-channel signal interference. Nevertheless, the high cost of imported mass cytometers has significantly hindered their widespread adoption in both scientific research and clinical diagnostics. In this study, the development of a domestically manufactured mass cytometer was reported, representing a significant advancement in high-end analytical instrumentation. The system integrated a pneumatic sample introduction module, an inductively coupled plasma (ICP) ion source, a quadrupole ion transmission system, a time-of-flight mass spectrometer (TOF MS), and a high-frequency data acquisition module. Performance evaluation was conducted using multi-element metal standard solutions. The instrument achieved a full mass range resolution exceeding 550 (full width at half maximum, FWHM). At a sample flow rate of 30 μL/min, the sensitivity for 159Tb+ reaches 9×105 cps/pg. Limits of detection are determined to be 0.4 ng/L for Cs, 0.2 ng/L for Tb, and 0.04 ng/L for Ir. An 8 h mass stability test demonstrates a mass drift of less than 0.1 u. Furthermore, the detection of cell surface protein expression using the developed instrument shows high concordance with results obtained from a leading commercial system (Helios). The successful construction and validation of this instrument fill a critical technological gap in the domestic market for high-performance mass cytometry. Its demonstrated resolution, sensitivity, stability, and analytical accuracy establish a robust platform for high-dimensional single-cell analysis. This achievement not only supports the expansion of multi-parameter cellular studies in basic research, but also holds substantial potential for translational applications in clinical diagnostics and precision medicine.

     

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