Single cell analysis in full body quartz glass chips with native UV laser-induced fluorescence detection
Single cell analysis in full body quartz glass chips with native UV laser-induced fluorescence detection
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DOI：
10.1016/j.chroma.2008.07.013
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发表时间：
2008-10-03
影响因子：
4.1
通讯作者：
Anselmetti, Dario
中科院分区：
文献类型：
--
作者：
Greif, Dominik;Galla, Lukas;Anselmetti, Dario
为了研究个体和非均匀的细胞反应，例如对外部刺激的反应，单细胞分析是强制性的，并且可以在蛋白质组学以及未来系统生物学的其他领域提供新的认知。在这里，我们报告了基于全身石英玻璃微流控芯片（QG芯片）的单细胞分析的新型芯片架构，该芯片扩展了我们以前在聚二甲基硅氧烷（PDMS）芯片中的研究，并提高了天然紫外激光诱导荧光（UV-LIF）检测的检测灵敏度。所有S/N比为11.9的10 nM色氨酸溶液的检测。其给出2.5 nM的理论检测限（S/N=3）。是可能的通过这些优化，将各自浓度为100 μ g/mL（等于4 μ M、0.4 μ M和2.2 μ M）的三种蛋白质α-胰凝乳蛋白酶原A、卵清蛋白和过氧化氢酶电动注射，并且可以以接近基线的分辨率分离。此外，荧光光谱（激发波长，λ（ex）= 266 nm）清楚地证明了与PDMS芯片和PDMS石英窗（PQW）芯片相比，QG芯片的有利性质，如非常高的UV透明度和几乎消失的背景荧光。最后，我们利用改进的灵敏度为单细胞电泳的草地贪夜蛾（Sf 9）昆虫细胞。(C)2008 Elsevier B. V.保留所有权利。
In order to investigate the individual and inhomogenous Cellular response, e.g. to external Stimuli, single cell analysis is mandatory and may provide new cognitions in proteomics as well as in other fields of systems biology in the future. Here, we report on novel chip architectures for single cell analysis based on full body quartz glass microfluidic chips (QG chips) that extend our previous studies in polydimethylsiloxane (PDMS) chips, and enhance the detection sensitivity of native UV laser-induced fluorescence (UV-LIF) detection. Detection of a 10 nM tryptophan Solution With ail S/N ratio of 11.9. which gives a theoretical limit of detection of 2.5 nM (with S/N=3). was possible. With these optimizations the three proteins alpha-chymotrypsinogen A, ovalbumin and catalase each at a concentration of 100 mu g/mL (equal to 4 mu M, 0.4 mu M and 2.2 mu M) were injected electrokinetically and could be separated with nearly baseline resolution. Furthermore, fluorescence spectra (excitation wavelength, lambda(ex) = 266 nm) clearly demonstrate the favourable properties like the very high UV transparency and the nearly vanishing background fluorescence of the QG chips as compared to PDMS chips and to PDMS quartz window (PQW) chips. Finally we exploit the improved sensitivity for single cell electropherograms of Spodoptera frugiperda (Sf9) insect cells. (C) 2008 Elsevier B.V. All rights reserved.