高等学校化学学报 ›› 2015, Vol. 36 ›› Issue (10): 1900.doi: 10.7503/cjcu20150282

• 分析化学 • 上一篇    下一篇

基于WST-8细胞活性实验的表面增强拉曼光谱研究

杨津1,2, 刘卓3, 王晓慧2, 隋会敏1, 毛竹1, 赵冰1()   

  1. 1. 吉林大学超分子结构与材料国家重点实验室, 长春 130012
    2. 吉林大学基础医学院, 3. 吉林大学中日联谊医院, 长春 130021
  • 收稿日期:2015-04-13 出版日期:2015-10-10 发布日期:2015-09-18
  • 作者简介:联系人简介: 赵冰, 男, 博士, 教授, 博士生导师, 主要从事光谱分析研究. E-mail:zhaob@jlu.edu.cn
  • 基金资助:
    国家自然科学基金(批准号: 21273091, 21221063, 21327803, 2141114023)和吉林省科技厅青年基金(批准号: 20130522181JH)资助

WST-8 Assay for Living Cell Viability Detection by Surface-enhanced Raman Spectroscopy

YANG Jin1,2, LIU Zhuo3, WANG Xiaohui2, SUI Huimin1, MAO Zhu1, ZHAO Bing1,*()   

  1. 1. State Key Laboratory of Supramolecular Structure and Materials, Jilin University, Changchun 130012, China
    2. College of Basic Medical Sciences, 3. China-Japan Union Hospital, Jilin University, Changchun 130021, China
  • Received:2015-04-13 Online:2015-10-10 Published:2015-09-18
  • Contact: ZHAO Bing E-mail:zhaob@jlu.edu.cn
  • Supported by:
    † Supported by the National Natural Science Foundation of China(Nos.21273091, 21221063, 21327803, 2141114023) and the Youth Fund of Science and Technology Department of Jilin Province, China(No.20130522181JH)

摘要:

采用常规的用于测定活细胞个数的试剂2-(2-甲氧基-4-硝苯基)-3-(4-硝苯基)-5-(2,4-二磺基苯)-2H-四唑单钠盐(WST-8四唑盐)建立了一种快速测定细胞活性的表面增强拉曼散射光谱(SERS)新方法. 考察了pH值及还原剂烟酰胺腺嘌噙二核苷酸(NADH)用量对WST-8甲瓒信号的影响. 结果表明, 与传统的WST-8比色法相比, SERS法具有更高的检测灵敏度和选择性, 对WST-8甲瓒的检测限可达到10 μmol/L.

关键词: 2-(2-甲氧基-4-硝苯基)-3-(4-硝苯基)-5-(2, 4-二磺基苯)-2H-四唑单钠盐(WST-8), 表面增强拉曼光谱, 细胞活性检测

Abstract:

Surface-enhanced Raman scattering(SERS) technique was used to develop and optimize a novel and quantitative 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium monosodium salt(WST-8) assay for living cell viability. The influences of pH and dosage of reductant nicoti-namide adenine dinucleotide(NADH) on WST-8 formazan signal were examined. SERS signals of formazan with high resolution and great diversity between WST-8 and formazan were observed in neutral conditions. Meanwhile, the signals of formazan(560, 1351, 1598, 1616 cm-1) could be found clearly when the concentration of NADH was four times that of WST-8. Compared with the traditional method, this assay appears higher sensitivity and selectivity for WST-8 formazan. Moreover, the detection limit of formazan is as low as 10 μmol/L.

Key words: 2-(2-Methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2, 4-disulfophenyl)-2H-tetrazolium monosodium salt(WST-8), Surface-enhanced Raman scattering(SERS), Cell viability detection

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