Shenzhen Key Laboratory for Minimal Invasive Medical Technologies, Institute of Optical Imaging and Sensing, Graduate School at Shenzhen, Tsinghua University, Shenzhen, 518055, Guangdong, China.
Department of Physics, Tsinghua University, Beijing, 100084, China.
Anal Bioanal Chem. 2020 May;412(12):2731-2741. doi: 10.1007/s00216-020-02528-5. Epub 2020 Mar 10.
A one-step synthesis using the reversed-phase suspension polymerization method and ultraviolet light curing is proposed for preparing the Raman-encoded suspension array (SA). The encoded microcarriers are prepared by doping the Raman reporter molecules into an aqueous phase, and then dispersing the aqueous phase in an oil phase and curing by ultraviolet light irradiation. The multiplexed biomolecule detection and various concentration experiments confirm the qualitative and quantitative analysis capabilities of the Raman-encoded SA with a limit of detection of 52.68 pM. The narrow bandwidth of the Raman spectrum can achieve a large number of codes in the available spectral range and the independence between the encoding channel and the fluorescent label channel provides the encoding method with high accuracy. This preparation method is simple and easy to operate, low in cost, and high in efficiency. A large number of hydrogel-based encoding microbeads could be quickly obtained with good biocompatibility. Most importantly, concentrating plenty of Raman reporter molecules inside the microbeads increases the signal intensity and means the molecular assembly is not limited by the functional groups; thus, the types of materials available for Raman encoding method are expanded. Furthermore, the signal intensity-related encoding method is verified by doping different proportions of Raman reporter molecules with our proposed synthesis method, which further increases the detection throughput of Raman-encoded SA. Graphical Abstract.
提出了一种采用反相悬浮聚合方法和紫外光固化一步合成法制备拉曼编码悬浮阵列(SA)的方法。编码微载体是通过将拉曼报告分子掺杂到水相中,然后将水相分散在油相中,并通过紫外光照射进行固化而制备的。通过多重生物分子检测和各种浓度实验,证实了拉曼编码 SA 的定性和定量分析能力,检测限低至 52.68 pM。拉曼光谱的窄带宽可以在可用光谱范围内实现大量编码,并且编码通道与荧光标记通道之间的独立性为编码方法提供了高精度。该制备方法简单易行,成本低,效率高。可以快速获得大量具有良好生物相容性的水凝胶基编码微球。最重要的是,将大量拉曼报告分子浓缩在微球内可以提高信号强度,并且意味着分子组装不受官能团的限制;因此,扩大了可用于拉曼编码方法的材料类型。此外,通过我们提出的合成方法掺杂不同比例的拉曼报告分子,对与信号强度相关的编码方法进行了验证,进一步提高了拉曼编码 SA 的检测通量。