Yan Songyuan, Rajestari Zarya, Morse Timothy Clifford, Li Harbour, Kulinsky Lawrence
Materials and Manufacturing Technology, University of California Irvine, 5200 Engineering Hall, Irvine, CA 92627, USA.
Mechanical and Aerospace Engineering, University of California Irvine, 5200 Engineering Hall, Irvine, CA 92627, USA.
Micromachines (Basel). 2024 Feb 29;15(3):341. doi: 10.3390/mi15030341.
The presented study demonstrates the capability of the template-based electrokinetic assembly (TEA) and guidance to manipulate and capture individual biological cells within a microfluidic platform. Specifically, dielectrophoretic (DEP) focusing of K-562 cells towards lithographically-defined "wells" on the microelectrodes and positioning singles cells withing these "wells" was demonstrated. K-562 lymphoblast cells, are widely used in immunology research. The DEP guidance, particularly involving positive DEP (pDEP), enables the controlled guidance and positioning of conductive and dielectric particles, including biological cells, opening new directions for the accurate and efficient microassembly of biological entities, which is crucial for single cell analysis and other applications in biotechnology. The investigation explores the use of glassy carbon and gold as electrode materials. It was established previously that undiluted physiological buffer is unsuitable for inducing positive DEP (pDEP); therefore, the change of media into a lower ionic concentration is necessary. After pDEP was observed, the cells are resubmerged in the Iscove's modified Dulbecco's medium (IMEM), a cell culturing media, and incubated. A dead/alive staining assay was performed on the cells to determine their survival in the diluted buffer for the period required to capture them. The staining assay confirmed the cells' survival after being immersed in the diluted biological buffer necessary for electrokinetic handling. The results indicate the promise of the proposed electrokinetic bio-sorting technology for applications in tissue engineering, lab-on-a-chip devices, and organ-on-a-chip models, as well as contributing to the advancement of single cell analysis.
本研究展示了基于模板的电动组装(TEA)在微流控平台内操纵和捕获单个生物细胞的能力及导向作用。具体而言,展示了将K-562细胞通过介电泳(DEP)聚焦到微电极上光刻定义的“阱”中,并将单个细胞定位在这些“阱”内。K-562淋巴母细胞广泛应用于免疫学研究。DEP导向,特别是涉及正介电泳(pDEP),能够对包括生物细胞在内的导电和介电颗粒进行可控导向和定位,为生物实体的精确高效微组装开辟了新方向,这对于单细胞分析和生物技术中的其他应用至关重要。该研究探索了使用玻璃碳和金作为电极材料。先前已确定未稀释的生理缓冲液不适用于诱导正介电泳(pDEP);因此,有必要将介质换成离子浓度较低的溶液。在观察到pDEP后,将细胞重新浸入Iscove改良的杜氏培养基(IMEM,一种细胞培养基)中并进行培养。对细胞进行死活染色测定,以确定它们在捕获所需时间段内在稀释缓冲液中的存活情况。染色测定证实了细胞在浸入电动处理所需的稀释生物缓冲液后仍能存活。结果表明,所提出的电动生物分选技术在组织工程、芯片实验室设备和芯片器官模型中的应用具有前景,也有助于单细胞分析的发展。