National Yang-Ming University, Institute of Biophotonics, Taipei, Taiwan.
J Biomed Opt. 2014 Jan;19(1):011012. doi: 10.1117/1.JBO.19.1.011012.
Cellular micropattering has been increasingly adopted in quantitative biological experiments. A Q-switched pulsed neodymium-doped yttrium ortho-vanadate (Nd∶YVO4) laser directed in-situ microfabrication technique for cell patterning is presented. A platform is designed uniquely to achieve laser ablation. The platform is comprised of thin gold coating over a glass surface that functions as a thermal transducer and is over-layered by a cell repellant polymer layer. Micropatterns are engraved on the platform, subsequently exposing specific cell adhesive micro-domains by ablating the gold-polymer coating photothermally. Experimental results indicate that the proposed approach is applicable under culture conditions, viable toward cells, and has a higher engraving speed. Possible uses in arraying isolated single cells on the platform are also shown. Additionally, based on those micro-patterns, dynamic cellular morphological changes and migrational speed in response to geometrical barriers are studied to demonstrate the potential applications of the proposed approach. Our results further demonstrate that cells in narrower geometry had elongated shapes and higher migrational speed than those in wider geometry. Importantly, the proposed approach will provide a valuable reference for efforts to study single cell dynamics and cellular migration related processes for areas such as cell division, wound healing, and cancer invasion.
细胞微图案化已越来越多地应用于定量生物学实验中。本研究提出了一种基于调 Q 脉冲掺钕钒酸钇(Nd∶YVO4)激光的原位微加工技术,用于细胞图案化。本研究设计了一个独特的平台来实现激光烧蚀。该平台由玻璃表面上的薄金涂层组成,充当热转换器,并覆盖有细胞排斥聚合物层。在平台上刻蚀微图案,随后通过光热烧蚀消融金-聚合物涂层,暴露特定的细胞黏附微区。实验结果表明,该方法适用于培养条件下,对细胞是可行的,并且具有更高的雕刻速度。还展示了在平台上排列分离的单细胞的可能用途。此外,基于这些微图案,研究了细胞对几何障碍的动态形态变化和迁移速度,以展示该方法的潜在应用。我们的结果进一步表明,在较窄的几何形状中,细胞具有更长的形状和更高的迁移速度,而在较宽的几何形状中则相反。重要的是,该方法将为研究细胞分裂、伤口愈合和癌症侵袭等领域的单细胞动力学和细胞迁移相关过程提供有价值的参考。