Spectral-optical-tweezer-assisted fluorescence multiplexing system for QDs-encoded bead-array bioassay
Spectral-optical-tweezer-assisted fluorescence multiplexing system for QDs-encoded bead-array bioassay
复制标题
用于量子点编码珠阵列生物测定的光谱光镊辅助荧光复用系统
DOI：
10.1016/j.bios.2019.01.004
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发表时间：
2019-03-15
影响因子：
12.6
通讯作者：
Cao, Donglin
中科院分区：
文献类型：
--
作者：
He, Qinghua;Chen, Xuejing;Cao, Donglin
作为生物分子多重检测的有效工具，微珠阵列可以实现对多个目标的免分离检测，适合于分析活体中有价值的和稀缺的样本，如抗原和抗体。在此，我们提出了一种光谱光钳辅助的荧光多路复用系统来分析生物分子共轭微珠阵列。使用光钳将珠子捕获并锁定在焦点处接受刺激，提供了稳定和优化的分析条件。移动系统焦点，扫描样品载玻片，经多路检测后，实现了对量子点编码微珠阵列的发射采集。用光谱仪采集和记录荧光发射光谱。通过识别解码峰的位置和统计发射光谱标签信号的强度，实现了对目标的定性和定量检测。作为概念验证研究，我们利用该系统对单个样品中的各种类型的抗-Ig G进行了多重检测，检测下限达到了1.52 pm，线性范围为0.31~10 nM。通过对实验条件的进一步优化，实现了夹心法对人血清中靶向免疫球蛋白的特异性检测，检测限低至0.23 pm，线性范围为0.88~28 pm，验证了该方法在实际样品中的实际应用。
As an efficient tool in the multiplexed detection of biomolecules, bead-array could achieve separation-free detection to multiple targets, making it suitable to analyze valuable and scarce samples like antigen and antibody from living organism. Herein, we propose a spectral-optical-tweezer-assisted fluorescence multiplexing system to analyze biomolecule-conjugated bead-array. Using optical tweezer, we trapped and locked beads at the focus to accept stimulation, offering a stable and optimized analysis condition. Moving the system focus and scanning the sample slide, we achieved emissions collection to QDs-encoded bead-array after the multiplexed detection. The emission spectra of fluorescence were collected and recorded by the spectrometer. By recognizing locations of decoding peaks and counting the intensities of label signals of emission spectra, we achieved qualitative and quantitative detection to targets. As proof-of-concept studies, we use this system to carry out multiplexed detection to various types of anti-IgG in the single sample and the detection limit reaches 1.52 pM with a linear range from 0.31 to 10 nM. Through further optimization of experimental conditions, we achieved specific detection to target IgG with sandwich method in human serum and the detection limit reaches as low as 0.23 pM with a linear range from 0.88 to 28 pM, validating the practical application of this method in real samples.