Mass Cytometry: Technique for Real Time Single Cell Multitarget Immunoassay Based on Inductively Coupled Plasma Time-of-Flight Mass Spectrometry
Mass Cytometry: Technique for Real Time Single Cell Multitarget Immunoassay Based on Inductively Coupled Plasma Time-of-Flight Mass Spectrometry
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DOI：
10.1021/ac901049w
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发表时间：
2009-08-15
影响因子：
7.4
通讯作者：
Tanner, Scott D.
中科院分区：
文献类型：
--
作者：
Bandura, Dmitry R.;Baranov, Vladimir I.;Tanner, Scott D.
描述了一种用于对单个生物细胞或其它微粒进行真实的实时分析的新型仪器。该仪器基于电感耦合等离子体飞行时间质谱法，并且包括三孔等离子体-真空接口、用于使离子与中性组分解耦的直流四极转向光学器件、区分低质量主导等离子体离子的rf四极离子导向器、点对平行聚焦de四极偶极子、正交加速反射器分析器、离散倍增极快速离子检测器，和8位1 GHz数字转换器。76.8 kHz的高光谱产生频率提供了从每个粒子引起的瞬态离子云收集多个光谱的能力，通常持续时间为200-300 μ s。它示出的瞬态可以单独地在每秒1100个粒子的峰值频率下被解析和表征。设计考虑和优化数据。仪器的品质因数是在标准电感耦合等离子体（ICP）操作条件下测量的（m/z = 159时为900，标准样品引入系统的灵敏度为> 1.4 × 10（8）离子计数/秒/mg L-1 Tb，丰度灵敏度为（6 x 10（-4））-（1.4 x 10（-3））（分别为后质量和前质量）。质量范围（m/z = 125-215）和丰度灵敏度足以用于具有多达60种不同可用元素标签的元素免疫测定。当500）时，其在（1.5 × 10（-3））-（5.0 × 10（-3））丰度灵敏度下提供> 2.4 × 10（8）cps/mg L-1的Tb。实时同时检测多个同位素从个别1.8 μ m聚苯乙烯头标记镧系元素。提供了用镧系元素标记的抗体免疫标记的模型细胞系和白血病患者样品的真实的单细胞20抗原表达测定。
A novel instrument for real time analysis of individual biological cells or other microparticles is described. The instrument is based on inductively coupled plasma time-of-flight mass spectrometry and comprises a three-aperture plasma-vacuum interface, a dc quadrupole turning optics for decoupling ions from neutral components, an rf quadrupole ion guide discriminating against low-mass dominant plasma ions, a point-to-parallel focusing de quadrupole doublet, an orthogonal acceleration reflectron analyzer, a discrete dynode fast ion detector, and an 8-bit 1 GHz digitizer. A high spectrum generation frequency of 76.8 kHz provides capability for collecting multiple spectra from each particle-induced transient ion cloud, typically of 200-300 mu s duration. It is shown that the transients can be resolved and characterized individually at a peak frequency of 1100 particles per second. Design considerations and optimization data are presented. The figures of merit of the instrument are measured under standard inductively coupled plasma (ICP) operating conditions ( 900 for m/z = 159, the sensitivity with a standard sample introduction system of > 1.4 x 10(8) ion counts per second per mg L-1 of Tb and an abundance sensitivity of (6 x 10(-4))-(1.4 x 10(-3)) (trailing and leading masses, respectively) are shown. The mass range (m/z = 125-215) and abundance sensitivity are sufficient for elemental immunoassay with up to 60 distinct available elemental tags. When 500) can be used, which provides > 2.4 x 10(8) cps per mg L-1 of Tb, at (1.5 x 10(-3))-(5.0 x 10(-3)) abundance sensitivity. The real-time simultaneous detection of multiple isotopes from individual 1.8 mu m polystyrene heads labeled with lanthanides is shown. A real time single cell 20 antigen expression assay of model cell lines and leukemia patient samples immuno-labeled with lanthanide-tagged antibodies is presented.