Microfluidic trapping of 3D spheres in microfluidic reaction systems for real time biosensing of flow chemistry derived compounds

第二个资助期的研究项目的主题将是基于3D有机物的多参数实时生物活性监测与微流控芯片实验室设计(第一资助期)的结合。将在高度环境控制下的在线实时生物分析用于流动下复杂的3D细胞聚集体的生物电子监测将是一项挑战。在第一个资助期，我们成功地建立了一种基于可行的2D细胞单层的快速、灵敏的实时分析工具，用于关于生物活性的化学合成产物/产品的阻抗监测。现在，芯片上的微流控捕获3D有机物，即再现体内情况的有机类型的心脏球体将是主要的研究课题。药物发现应在微流控芯片上实现，该芯片由两个层次组成：(I)用于微合成区域、微FFE区域和在(Ii)第二层次上以多孔形式连接3D微腔-电极的微通道，包括用于3D培养的定位通道。新型微腔阵列结构(200-500微米长，130-330微米深)，每个微腔至少有四个电极，将使用选择性激光刻蚀的创新技术在熔融二氧化硅衬底上生产。这些集成在多级微流控芯片上的微腔结构将通过混合多路复用器电子板进行接触，该混合多路复用器电路板是在第一个资助期间开发的阻抗多路复用器电路板的基础上开发的。对于生物靶标的同步多模测量，将在电子板上扩展一个用于场电位记录的电子模块。扩展的多模式系统包括在流体条件下的3D心簇的阻抗频谱、场电位记录和光子监测之间的切换，其允许实时分析生理、生物力学和电生理特性。因此，从第一个资助期开始，用于活细胞单层的微流控芯片将被一个新的模块扩展，用于在微腔阵列中捕获3D有机物和实时混合检测3D球体，这是基于过去证明的经验。

The topic of the research project of the 2nd funding period will be the combination of a 3D organoid based multiparametric real time biological activity monitoring and the adaptation of the microfluidic lab-on-chip design (1st funding period). Coupling an inline real time bioanalysis under a high degree of environmental control for the bioelectronic monitoring of complex 3D cell aggregates under flow will be the challenge. During the 1st funding period we were successful in establishing a fast, sensitive real time analytical tool based on viable 2D cell monolayer for impedimetric monitoring of chemical synthesis educts/products with regard to biological activity. Now the on-chip microfluidic trapping of 3D organoids i.e. organotypic cardiac spheres that recapitulate the in vivo situation will be the main research topic. The drug discovery shall be realized on a microfluidic chip consisting of two levels (i) for the microsynthesis area, the µFFE field and the microchannels connecting the 3D microcavity-electrode as a multi-well format on the (ii) second level including positioning channels for the 3D cultures. The novel microcavity array structures (200 – 500 µm length, 130-330 µm depth) with at least four electrodes per microcavity will be produced in fused silica substrates using the innovative technology of selective laser etching. These integrated microcavity structures on the multilevel microfluidic chip will be contacted by a hybrid multiplexer electronic board, which is based on the impedance multiplexer board developed during the 1st funding period. For the synchronous multimodal measurement of the biological targets, the electronic board will be extended by an electronic module for the field potential recording. The extended multimodal system comprises, a switch between impedance spectroscopy, field potential recording, and photonic monitoring of 3D cardiac clusters under fluidic conditions, which allows to analyse physiological biomechanical and electrophysiological properties in real-time. Thus, the microfluidic chip for viable cell monolayer from the 1st funding period will be extended by a novel module for 3D organoid trapping and real time hybrid live-sensing of 3D spheres in microcavity arrays based on experiences demonstrated in the past.