具有精密微纳结构的高通量编码微球库的建立及其编码性能的构效关系研究

In order to eliminate the encoding capacity limitations of encoded beads in the area of high-throughput multiplexed detections, construction of a precise micro/nanostructured barcode library with a large encoding capacity via a novel structure-fluorescence joint encoding strategy is proposed in this project. Moreover, the structure-activity relationship between assembled structure of microspheres and encoding property will also be investigated. By employing mesoporous beads as microcarriers, the precise structures are designed for their different regions. Firstly, specific scattered light as the first dimensional encoding will be produced by regulating the inner structure (including framework component and diameter of mesopores) of beads. Next, organic dyes are loaded inside mesopores to realize the second dimensional fluorescent encoding. Then, both magnetic nanoparticles and fluorescent quantum dots are assembled onto the outer surface to endow the microspheres with superparamagnetic and the third dimensional fluorescent encoding properties. The strategy of multi-dimensional structure-fluorescence joint encoding method will significantly enlarge the encoding capacity of barcodes through the precise design of micro/nanostructures. The final constructed barcode library could obtain three encoding information simultaneously through a single excitation laser, and it’s also a multiplexed encoding platform with a high encoding capacity and a magnetic manipulable property. This platform is promising in high-throughput multiplexed detection area of gene analysis, clinical diagnosis, drug screening, environmental monitoring, and even in the field of encryption and anti-counterfeiting.

针对编码微球在高通量多指标检测中的编码容量受限问题，本课题提出建立一种具有精密微纳结构、利用结构和荧光联合编码的高通量编码微球库，并研究微球组装结构与编码性能之间的构效关系。以介孔微球作为载体基质，对其进行了分区域的精密结构设计：首先通过调控微球的内部结构（包括骨架成分和介孔孔径大小），产生特异性的散射光作为第一维度的编码；其次利用介孔孔道装载荧光染料进行第二维度的荧光编码；再利用微球的外表面组装磁性纳米颗粒和荧光量子点，赋予微球超顺磁性和第三维度的荧光编码。通过精密的微纳结构设计，利用微球的结构和两种荧光元素进行多维度的联合编码，可显著扩充载体的编码容量。最终所构建的编码微球库是一种单色激光可同时激发三种编码要素、具有高通量编码能力和可磁场操纵的多指标编码平台，有望在基因分析、临床诊断、药物筛选、环境监测等高通量的多指标检测以及加密防伪等领域得到实际应用。

随着悬浮芯片技术的快速发展，具有高编码容量特性的核心部件-即基于微珠的编码微球，在单管多指标检测面临超高通量的应用需求时显得极为重要，目前的技术略显不足。针对上述本领域存在的瓶颈问题，我们首次创新地提出了“结构-荧光”联合编码策略，以求建立一个具有超高容量的编码微球库。基于我们首次揭示的介孔微球内部结构参数（例如孔隙率和基质成分）可改变微球在流式解码系统上的散射光强度，即发现了新的结构编码元素，再联合另外两种荧光编码元素-即绿色的异硫氰酸荧光素和红色的量子点，即可成功实现编码容量的显著扩充。该编码微球系统具有精密的微纳结构设计，其中异硫氰酸荧光素染料分子被封装于介孔孔道内部，而磁性纳米颗粒和量子点则被依次组装于介孔载体微球的外表面，从而可同时赋予微球两种荧光编码信息以及超顺磁性。最终，通过结构和荧光的联合编码组合，可成功实现单色激光激发的300重超高通量编码微球库的建立，这是目前已有报道的最高记录。此外，我们还对精密设计的编码微球系统进行了耐受稳定性和储存稳定性的评估，并证明了该平台在实际应用过程中的可行性和有效性。总之，本项目的研究成果说明我们所构建的编码微球系统对于超高通量的单管多指标检测是一种强大有效的工具，展现了良好的潜在应用价值。