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.
The plant cell mechanics, including turgor pressure and wall mechanical properties, not only determine the growth of plant cells, but also reflect the functional and structural changes of plant cells under biotic and abiotic stresses. However, there are currently no appropriate techniques allowing to monitor the complex mechanical properties of living plant cells non-invasively and continuously. In this work, quartz crystal microbalance with dissipation (QCM-D) monitoring technique with overtones (3–9) was used for the dynamic monitoring of adhesions of living tobacco BY-2 cells onto positively charged N,N-dimethyl-N-propenyl-2-propen-1-aminiumchloride homopolymer (PDADMAC)/SiO2 QCM crystals under different concentrations of mannitol (CM) and the subsequent effects of osmotic stresses. The cell viscoelastic index (CVIn) (CVIn = ΔD⋅n/ΔF) was used to characterize the viscoelastic properties of BY-2 cells under different osmotic conditions. Our results indicated that lower overtones of QCM could detect both the cell wall and cytoskeleton structures allowing the detection of plasmolysis phenomena; whereas higher overtones could only detect the cell wall’s mechanical properties. The QCM results were further discussed with the morphological changes of the BY-2 cells by an optical microscopy. The dynamic changes of cell’s generated forces or cellular structures of plant cells caused by external stimuli (or stresses) can be traced by non-destructive and dynamic monitoring of cells’ viscoelasticity, which provides a new way for the characterization and study of plant cells. QCM-D could map viscoelastic properties of different cellular structures in living cells and could be used as a new tool to test the mechanical properties of plant cells.
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