高灵敏、宽动态范围纳米流式检测装置的研制及应用

引用本文: 毛翠萍, 林垚, 牛倩, 薛乘风, 颜晓梅. 高灵敏、宽动态范围纳米流式检测装置的研制及应用[J]. 分析化学, 2020, 48(7): 838-846. doi: 10.19756/j.issn.0253-3820.201227 shu

Citation: MAO Cui-Ping, LIN Yao, NIU Qian, XUE Cheng-Feng, YAN Xiao-Mei. Development and Applications of A Highly Sensitive and Wide Dynamic Range Nano-Flow Cytometer[J]. Chinese Journal of Analytical Chemistry, 2020, 48(7): 838-846. doi: 10.19756/j.issn.0253-3820.201227 shu

高灵敏、宽动态范围纳米流式检测装置的研制及应用

厦门大学化学化工学院化学生物学系, 谱学分析与仪器教育部重点实验室, 化学生物学福建省重点实验室, 厦门 361005
基金项目:
本文系国家自然科学基金项目（Nos.21627811，21934004）资助

摘要: 基于纳米颗粒固有的多分散性，对其粒径及分布快速地进行高分辨表征是纳米颗粒相关研究和产品开发所面临的重大挑战。与电子显微镜、原子力显微镜、动态光散射、纳米颗粒追踪分析等常规使用的纳米颗粒表征技术相比，流式细胞术具有可单颗粒检测、快速、多参数、统计精确性高等优势。然而，由于检测灵敏度的局限性，商品化流式细胞仪难以检测粒径小于200 nm的聚苯乙烯纳米颗粒。结合瑞利散射和鞘流单分子荧光检测技术，本研究组研制了纳米流式检测装置（Nano-flow cytometer，nFCM），采用单光子计数雪崩光电二极管（Single photon counting avalanche photodiode，APD）作为检测器，低折射率二氧化硅纳米颗粒（SiO₂ NPs）的检测下限可达到24 nm，可基线分辨47、59、74、94和123 nm的SiO₂ NPs混合样本。由于nFCM散射光检测动态范围受限于单光子探测器的最大光子计数值，而纳米颗粒的散射光强度随粒径的增大呈指数上升，为实现纳米颗粒更全面的粒径分布检测，仪器的检测动态范围有待提升。本研究采用激光光束整形、双光阑降低背景信号、高量子效率光电倍增管检测器等策略对仪器进行多方位改进，以研制高灵敏、宽动态范围的nFCM。改进后的nFCM可实现直径59 nm SiO₂ NPs的高信噪比检测（S/N=119），可基线分辨59、74、94、123、160、180和222 nm的SiO₂ NPs混合样本，相比以APD为检测器的nFCM，粒径检测范围扩展约100 nm，并成功用于鼠伤寒沙门氏菌外膜囊泡（Outer membrane vesicles，OMVs）粒径分布的快速测定。

关键词:

English

Development and Applications of A Highly Sensitive and Wide Dynamic Range Nano-Flow Cytometer

Department of Chemical Biology, The MOE Key Laboratory of Spectrochemical Analysis&Instrumentation, The Key Laboratory for Chemical Biology of Fujian Province, Collaborative Innovation Center of Chemistry for Energy Materials, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China
Received Date: 24 April 2020
Revised Date: 14 May 2020
Fund Project:
This work was supported by the National Natural Science Foundation of China (Nos. 21627811, 21934004).

Abstract: Nanoparticles (NPs) are playing increasingly important roles in the fields of biomedicine, biochemical analysis, and energy materials. Because NPs are intrinsically heterogeneous, rapid and high resolution size distribution measurement of NPs is a challenging task. Compared with traditional approaches for the analysis of NPs, such as electron microscopy, atomic force microscopy, dynamic light scattering, and nanoparticle tracking analysis, flow cytometry is a well-established technique for high-throughput, quantitative, and multi-parameter analysis of individual cells and microscopic particles. However, it has been extremely difficult for conventional flow cytometry to detect polystyrene NPs smaller than 200 nm in diameter via light scattering. Employing strategies for single molecule fluorescence detection in a sheathed flow, we have recently developed Nano-flow cytometer (nFCM) that enables light scattering detection of single silica nanoparticles (SiO₂ NPs) as small as 24 nm in diameter. However, the dynamic range of the nFCM is limited by the maximum photon count of the single photon counting avalanche photodiode (APD) detector. Owing to the exponential dependence of scattered light intensity on particle size, there is an urgent need to enhance the dynamic range of the nFCM for the analysis of samples with wide size distributions. Adopting strategies of beam shaping with binary optical element, background reduction upon the combined usage of an iris-diaphragm and a rectangular aperture, and signal enhancement using high quantum efficiency PMT, here we reported the development of a highly sensitive and wide dynamic range nFCM. Sensitive light-scattering detection of single SiO₂ NPs as small as 59 nm in diameter was achieved with a signal-to-noise ratio of 119. Baseline separation of seven different sizes of SiO₂ NPs ranging from 59 to 222 nm was obtained. The as-developed nFCM was successfully applied to the size distribution analysis of outer membrane vesicles derived from Salmonella Typhimurium.

Key words:

1. [1]
  Zheng K Y, Setyawati M I, Leong D T, Xie J P. Coord. Chem. Rev.,2018,357:1-17
2. [2]
  Barile L, Vassalli G. Pharmacol. Ther.,2017,174:63-78
3. [3]
  Dadfar S M, Roemhild K, Drude N I, Stillfried S V, Knuchel R, Kiessling F, Lammers T. Adv. Drug Deliver. Rev.,2019,138:302-325
4. [4]
  Beik J, Khateri M, Khosravi Z, Kamrava S K, Kooranifar S, Ghaznavi H, Shakeri-Zadeh A. Coord. Chem. Rev.,2019,387:299-324
5. [5]
  Wu Z P, Wang Y L, Liu X B, Lv C, Li Y S, Wei D, Liu Z F. Adv. Mater.,2019,31(9):e1800716
6. [6]
  Li T T, Shi S X, Goel S, Shen X, Xie X X, Chen Z Y, Zhang H X, Li S, Qin X, Yang H, Wu C H, Liu Y Y. Acta Biomater.,2019,89:1-13
7. [7]
  Jin W, Maduraiveeran G. Trends Environ. Anal.,2017,14:28-36
8. [8]
  Bai X, Wang S Q, Yan X L, Zhou H Y, Zhan J H, Liu S, Sharma V K, Jiang G B, Zhu H, Yan B. ACS Nano,2020,14(1):289-302
9. [9]
  Homaeigohar S, Kabir R, Elbahri M. Sci. Rep.,2020,10:5191
10. [10]
  Zhou J J, Chizhik A I, Chu S, Jin D Y. Nature,2020,579(7797):41-50
11. [11]
  Vanderpol E, Coumans F A W, Grootemaat A E, Gardiner C, Sargent I L, Harrison P, Sturk A, van Leeuwen T G, Nieuwland R. J. Thromb. Haemost.,2014,12(7):1182-1192
12. [12]
  Shapiro H M. Practical Flow Cytometry; Hoboken:John Wiley & Sons, Inc.,2003
13. [13]
  Zhu S B, Ma L, Wang S, Chen C X, Zhang W Q, Yang L L, Hang W, Nolan J P, Wu L N, Yan X M. ACS Nano,2014,8(10):10998-11006
14. [14]
  Zhu S B, Yang L L, Long Y, Gao M, Huang T X, Hang W, Yan X M. J. Am. Chem. Soc.,2010,132(35):12176-12178
15. [15]
  Yang L L, Zhu S B, Hang W, Wu L, Yan X M. Anal. Chem.,2009,81(7):2555-2563
16. [16]
  Zhang S Y, Zhu S B, Yang L L, Zheng Y, Gao M, Wang S, Zeng J Z, Yan X M. Anal. Chem.,2012,84(15):6421-6428
17. [17]
  Ma L, Zhu S B, Tian Y, Zhang W Q, Wang S, Chen C X, Wu L N, Yan X M. Angew. Chem. Int. Edit.,2016,55(35):10239-10243
18. [18]
  Chen C C, Zhu S B, Wang S, Zhang W Q, Cheng Y, Yan X M. ACS Appl. Mater. Interfaces,2017,9(16):13913-13919
19. [19]
  Tian Y, Ma L, Gong M F, Su G Q, Zhu S B, Zhang W Q, Wang S, Li Z B, Chen C C, Li L H, Wu L N, Yan X M. ACS Nano,2018,12(1):671-680
20. [20]
  Lawrence W G, Varadi G, Entine G, Podniesinski E, Wallace P K. Proc. SPIE,2008,6859:68590M
21. [21]
  Zhao J J, You Z. Biomicrofluidics,2016,10(5):054111
22. [22]
  Shapiro H M. In Current Protocols in Cytometry:John Wiley & Sons, Inc.,2001:1.6.1-1.6.5
23. [23]
  Perfetto S P, Chattopadhyay P K, Wood J, Nguyen R, Ambrozak D, Hill J P, Roederer M. Cytometry A,2014,85(12):1037-1048
24. [24]
  Zhang W Q, Tian Y, Hu X X, He S B, Niu Q, Chen C, Zhu S B, Yan X M. Anal. Chem.,2018,90(21):12768-12775
25. [25]
  The Micro-Plus Flow Cytometer, Apogee Flow Systems, http://www.apogeeflow.com/products.php
26. [26]
  Roier S, Zingl F G, Cakar F, Durakovic S, Kohl P, Eichmann T O, Klug L, Gadermaier B, Weinzerl K, Prassl R, Lass A, Daum G, Reidl J, Feldman M F, Schild S. Nat. Commun.,2016,7:710515
27. [27]
  Schulz E, Goes A, Garcia R, Panter F, Koch M, Muller R, Fuhrmann K, Fuhrmann G. J. Control Release,2018,290:46-55
28. [28]
  Berleman J, Auer M. Environ. Microbiol.,2013,15(2):347-354
29. [29]
  Toyofuku M, Nomura N, Eberl L. Nat. Rev. Microbiol.,2019,17(1):13-24

扫一扫看文章

计量

PDF下载量: 4
文章访问数: 399
HTML全文浏览量: 13

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

高灵敏、宽动态范围纳米流式检测装置的研制及应用

English

Development and Applications of A Highly Sensitive and Wide Dynamic Range Nano-Flow Cytometer

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

高灵敏、宽动态范围纳米流式检测装置的研制及应用

English

Development and Applications of A Highly Sensitive and Wide Dynamic Range Nano-Flow Cytometer

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs