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使用高效细胞引入系统通过时间分辨电感耦合等离子体质谱法对单个酵母细胞(酿酒酵母)进行高灵敏度元素分析。

High sensitive elemental analysis of single yeast cells (Saccharomyces cerevisiae) by time-resolved inductively-coupled plasma mass spectrometry using a high efficiency cell introduction system.

作者信息

Groombridge Alexander S, Miyashita Shin-ichi, Fujii Shin-ichiro, Nagasawa Keisuke, Okahashi Tetsuya, Ohata Masaki, Umemura Tomonari, Takatsu Akiko, Inagaki Kazumi, Chiba Koichi

机构信息

Environmental Standards Section, National Metrology Institute of Japan, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan.

出版信息

Anal Sci. 2013;29(6):597-603. doi: 10.2116/analsci.29.597.

DOI:10.2116/analsci.29.597
PMID:23749124
Abstract

Trace elemental analysis of single yeast cells with time-resolved inductively coupled plasma mass spectrometry (ICP-MS) was successfully carried out, where a high efficiency cell introduction system (HECIS) consisting of the high performance concentric nebulizer (HPCN) and a low-volume (15 mL) on-axis spray chamber utilizing a sheath gas flow were used. Cell adsorption to the flow injector and sample tubing was reduced with the addition of a simple 4.3 mmol L(-1) of NaCl solution to the cell suspension and cell flowing liquid, allowing consecutive measurements without fear of significant contamination from previous measurements. Initially using a quadrupole mass analyzer ICP-MS (ICP-QMS) at its lowest integration time (10 ms), current spikes corresponding to separate cell events were detected for several elements (Mg, P, Ca, Mn, Fe, Cu, and Zn) on the introduction of the cell suspension. On comparing the number of peaks in the spectrum for phosphorous with the cell count using a haemocytometer, a reproducible cell transport efficiency of 75.0 ± 4.7% was achieved. Preliminary experiments into using time of flight ICP-MS (ICP-TOFMS) for single-cell analysis were carried out, allowing quasi-simultaneous multielement detection. The spectra of Mg, P, Ca, Mn, Fe, Cu, and Zn, with a time resolution of 1 ms were simultaneously obtained in one measurement. A relatively strong correlation was observed for the spectra between P and Zn (correlation factor 0.69), P and Mg (0.63), and Mg and Zn (0.63). These results indicate that the time resolved quasi-simultaneous multielement measurement may be useful for the correlation analysis of multielements in cells.

摘要

采用时间分辨电感耦合等离子体质谱法(ICP-MS)成功地对单个酵母细胞进行了痕量元素分析,其中使用了由高性能同心雾化器(HPCN)和利用鞘气流动的小体积(15 mL)同轴喷雾室组成的高效细胞引入系统(HECIS)。通过向细胞悬浮液和细胞流动液中添加简单的4.3 mmol L(-1) NaCl溶液,减少了细胞在流动注射仪和样品管上的吸附,从而能够进行连续测量,而无需担心先前测量造成的显著污染。最初在最低积分时间(10 ms)下使用四极杆质谱分析仪ICP-MS(ICP-QMS),在引入细胞悬浮液时,检测到了几种元素(Mg、P、Ca、Mn、Fe、Cu和Zn)对应于单个细胞事件的电流尖峰。通过使用血细胞计数器将磷光谱中的峰数与细胞计数进行比较,实现了75.0±4.7%的可重复细胞传输效率。开展了使用飞行时间ICP-MS(ICP-TOFMS)进行单细胞分析的初步实验,实现了准同时多元素检测。在一次测量中同时获得了时间分辨率为1 ms的Mg、P、Ca、Mn、Fe、Cu和Zn的光谱。观察到P与Zn(相关系数0.69)、P与Mg(0.63)以及Mg与Zn(0.63)的光谱之间存在相对较强的相关性。这些结果表明,时间分辨准同时多元素测量可能有助于细胞中多元素的相关性分析。

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