基于高灵敏腔体微电极阵列的抗肿瘤药物筛选新型传感器

In-vitro screening for anticancer drug is important for cancer treatment and drug discovery, and the microelectrode-based impedance biosensor gives a very promising method for real-time, dynamic, continuous activity monitoring. However, these biosensors have some disadvantages, such as low sensitivity and anti-interferential capability, which is due to uncontrolled resistance gradients and charge density. To this end, a sensing device will be developed using microelectrode arrays, coupled with nanomaterials technology and surface plasma resonance. We will focus on the development of 3D arrays constructed by gold nanoparticles and mesoporous materials, which exhibits surface plasmon polaritons that can be applied to control charge density. The resistance gradient will be adjusted by template parameters and gel-layer thickness. Also, the sensing performance and mechanism will be explored in terms of sensitivity, anti-interference and biocompatibility. Furthermore, breast cancer cells will be used as a model to investigate its practical applications of sensors. These results will allow for the development of new theory and methodology of biosensors for anticancer drug screening. As such, this study will provide information to better evaluate modularized microelectrode arrays for drug screening in the future. This study will be of great importance in the development of high-throughput biosensors for in-vitro anticancer drug screening.

抗肿瘤药物体外筛选对癌症治疗和药物研究极为重要，基于微纳电极阵列的阻抗传感器因能实时、动态、持续地反应细胞活性状态，有望实现抗肿瘤药物高通量筛选。但现有阻抗传感器由于电阻梯度和电荷密度不可控，在结构和性能上造成抗干扰能力差和灵敏度低。针对这一问题，本项目将微纳电极阵列、纳米材料修饰、等离子谐振等技术与传感器结合，开展基于高灵敏腔体微电极阵列的抗肿瘤药物筛选新型传感器研究。重点研制一种具有纳米结构的三维腔体微电极阵列传感器，通过调节模板参数和凝胶层厚度对腔体电阻梯度实现有效控制，结合表面等离激元效应来调控电极表面电荷密度；研究该传感器的检测性能及传感机理，并以乳腺癌细胞为检测模型来探索其用于细胞活性检测及药物筛选的可行性，进而建立用于抗肿瘤药物筛选的传感器新理论和新方法，为未来研究模块化药物筛选微电极阵列提供传感器基础和技术支撑，对实现高通量抗肿瘤药物体外筛选具有重要的科学意义和应用价值。

抗肿瘤药物体外筛选对疾病治疗和药物研究极为重要，基于微纳电极阵列的阻抗传感器因能实时、动态、持续地反应细胞活性状态，有望实现抗肿瘤药物高通量筛选。但现有阻抗传感器由于电阻梯度和电荷密度不可控，在结构和性能上造成抗干扰能力差和灵敏度低。针对这一问题，本研究将微纳电极阵列、纳米材料修饰、等离子谐振等技术与生物传感器结合，开展基于高灵敏度腔体微电极阵列的抗肿瘤药物筛选新型传感器研究。在ITO玻璃表面大面积制作有序PS微球单分散薄膜，结合单原子层沉积法（ALD）和电感耦合等离子体刻蚀法（ICP）制备具有电阻梯度的有序多孔Al2O3/ITO三维腔体微电极阵列，通过调节模板参数和球腔厚度，实现对传感器腔体电阻梯度的有效控制。依据电极表面电荷密度可控设计准则，将CdO/ZnO和GaInPSb纳米晶组装于球腔表面，结合表面等离激元效应调控电极表面电荷密度。选择一定浓度的、经胰蛋白酶与超声处理的细菌细胞（红色荧光染色剂标记金黄葡萄球菌、黄色荧光蛋白标记志贺氏短杆菌、绿色荧光蛋白标记炭疽杆菌芽孢）分别放置球腔内进行预实验，研究不同状态同种细菌细胞、不同形状多种细菌细胞、不同尺寸细菌细胞在球腔阵列中的排列情况。在载有数百万细菌细胞的球腔阵列基底上，借助荧光标记、激光共聚焦扫描显微镜和扫描电子显微镜，实现了单个细菌细胞高效定位，形成细胞阻抗传感器。将乳腺癌细胞放置球腔中作为生物敏感元件，探索其用于细胞活性检测及药物筛选的可行性，通过测量细胞阻抗变化，获得阿霉素与细胞作用的动态信息，将微电极阵列、表面等离激元调控、新型石墨烯场效应生物传感器等技术集成，较传统液栅结构灵敏度提高20%，检测阿霉素引起细胞阻抗变化的分辨率达10nM左右。重点研究了该传感器的检测性能及传感机理，进而建立用于抗肿瘤药物筛选传感器的新理论和新方法，为未来研究模块化药物筛选微电极阵列提供传感器基础和技术支撑，对实现高通量抗肿瘤药物体外筛选具有重要的科学意义和应用价值。