Shen Haibo, Zhou Tiean, Hu Jiajin
Cell Mechanics and Biosensing Institute, Hunan Agricultural University, 405 Life Sciences Building, Furong District, Changsha, Hunan, 410128, China.
College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha, Hunan, 410128, China.
Anal Bioanal Chem. 2017 Nov;409(27):6463-6473. doi: 10.1007/s00216-017-0591-4. Epub 2017 Sep 9.
In this study, we present a novel design of interference-free, negligible installation-induced stress, suitable for the fabrication of high-throughput quartz crystal microbalance (HQCM) chips. This novel HQCM chip configuration was fabricated using eight independent yet same-batch quartz crystal resonators within a common glass substrate with eight through-holes of diameter slightly larger than that of the quartz resonator. Each quartz resonator's rim was adhered to the inner part of the through-hole via silicone glue to form the rigid (quartz)-soft (silicone)-rigid (glass) structure (RSRS) which effectively eliminates the acoustic couplings among different resonators and largely alleviates the installation-induced stresses. The consistence of the eight resonators was verified by very similar equivalent circuit parameters and very close response slopes to liquid density and viscosity. The HQCM chip was then employed for real-time and continuous monitoring of H9C2 cardiomyoblast adhesions and viscoelastic changes induced by the treatments of two types of drugs: drugs that affect the cytoskeletons, including nocodazole, paclitaxel, and Y-27632, and drugs that affect the contractile properties of the cells: verapamil and different dosages of isoprenaline. Meanwhile, we compared the cytoskeleton affecting drug-induced viscoelastic changes of H9C2 with those of human umbilical vein endothelial cells (HUVECs). The results described here provide the first solution to fabricate HQCM chips that are free from the limitation of resonator number, installation-induced stress, and acoustic interferences among resonators, which should find wide applications in areas of cell phenotype assay, cytotoxicity test, drug evaluation and screening, etc. Graphical abstract Schematic illustration of the principle and configuration of interference-free high-throughput QCM chip to evaluate and screen drugs based on cell viscoelasticity.
在本研究中，我们提出了一种新型的无干扰、安装引起的应力可忽略不计的设计，适用于高通量石英晶体微天平（HQCM）芯片的制造。这种新型的HQCM芯片结构是在一个普通玻璃基板内使用八个独立但同批次的石英晶体谐振器制造而成的，该玻璃基板有八个直径略大于石英谐振器的通孔。每个石英谐振器的边缘通过硅胶胶水粘附到通孔的内部，形成刚性（石英）-柔性（硅胶）-刚性（玻璃）结构（RSRS），有效地消除了不同谐振器之间的声学耦合，并在很大程度上减轻了安装引起的应力。通过非常相似的等效电路参数以及对液体密度和粘度非常接近的响应斜率，验证了这八个谐振器的一致性。然后，将HQCM芯片用于实时连续监测H9C2心肌成纤维细胞的粘附情况以及由两种药物处理引起的粘弹性变化：影响细胞骨架的药物，包括诺考达唑、紫杉醇和Y-27632，以及影响细胞收缩特性的药物：维拉帕米和不同剂量的异丙肾上腺素。同时，我们比较了影响细胞骨架的药物诱导的H9C2与人类脐静脉内皮细胞（HUVECs）的粘弹性变化。这里描述的结果提供了制造HQCM芯片的首个解决方案，该芯片不受谐振器数量、安装引起的应力和谐振器之间声学干扰的限制，应在细胞表型分析、细胞毒性测试、药物评估和筛选等领域得到广泛应用。图形摘要 基于细胞粘弹性评估和筛选药物的无干扰高通量QCM芯片的原理和结构示意图。
