Continuous monitoring of bisulfide variation in microdialysis effluents by on-line droplet-based microfluidic fluorescent sensor.
Continuous monitoring of bisulfide variation in microdialysis effluents by on-line droplet-based microfluidic fluorescent sensor.
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DOI：
10.1016/j.bios.2013.12.056
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发表时间：
2014-05
影响因子：
12.6
通讯作者：
Xiaocui Zhu;Lei Xu;Tongbo Wu;Anqin Xu;Meiping Zhao;Shaorong Liu
中科院分区：
文献类型：
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作者：
Xiaocui Zhu;Lei Xu;Tongbo Wu;Anqin Xu;Meiping Zhao;Shaorong Liu
我们展示了一种新型的荧光传感器，通过使用纳米颗粒谷胱甘肽-异硫氰酸荧光素（AuNP-GSH-FITC）探针耦合在线液滴为基础的微流控芯片的变化，实时和连续监测微透析流出液中的二硫化物。首次研制了AuNP-GSH-FITC荧光探针，并将其应用于荧光定量PCR检测体系中的二硫键，其线性范围为0.1 μM ~ 5.0 μM，检测限约为50 nM。响应时间小于2 min。借助于共固定的巯基-聚乙二醇，探针在高盐度溶液中表现出优异的稳定性和重现性，包括人工脑脊液（aCSF）。通过向探针溶液中加入0.1%乙二醛，该测定允许在微摩尔浓度水平下在半胱氨酸存在下定量二硫化物。利用AuNP-GSH-FITC探针，进一步构建了基于液滴的微流控荧光传感器，用于在线监测微透析流出液中二硫键的变化。以荧光显微镜-电荷耦合器件相机为检测器，微透析/微流控芯片集成装置对样品中二硫化物的检测限为2.0 μM，线性范围为5.0 ~ 50 μM。该方法已成功应用于aCSF和血清样品中二硫化物变化的在线测量。这将是一个非常有用的工具，用于跟踪细胞外液中的二硫化物或硫化氢的变化。
We demonstrate a novel fluorescent sensor for real-time and continuous monitoring of the variation of bisulfide in microdialysis effluents by using a nanoparticle–glutathione-fluorescein isothiocyanate (AuNP–GSH-FITC) probe coupled with on-line droplet-based microfluidic chip. The AuNP–GSH-FITC fluorescent probe was firstly developed and used for bisulfide detection in bulk solution by quantitative real-time PCR, which achieved a linear working range from 0.1 μM to 5.0 μM and a limit of detection of ~50 nM. The response time was less than 2 min. With the aid of co-immobilized thiol-polyethylene glycol, the probe exhibited excellent stability and reproducibility in high salinity solutions, including artificial cerebrospinal fluids (aCSF). By adding 0.1% glyoxal to the probe solution, the assay allowed quantification of bisulfide in the presence of cysteine at the micro-molarity level. Using the AuNP–GSH-FITC probe, a droplet-based microfluidic fluorescent sensor was further constructed for online monitoring of bisulfide variation in the effluent of microdialysis. By using fluorescence microscope-charge-coupled device camera as the detector, the integrated microdialysis/microfluidic chip device achieved a detection limit of 2.0 μM and a linear response from 5.0 μM to 50 μM for bisulfide in the tested sample. The method was successfully applied for the on-line measurement of bisulfide variation in aCSF and serum samples. It will be a very useful tool for tracking the variation of bisulfide or hydrogen sulfide in extracellular fluids.