Sugiura Seiryo, Nishimura Satoshi, Yasuda Soichiro, Hosoya Yumiko, Katoh Kaoru
Computational Biomechanics Laboratory, Department of Human and Engineered Environmental Studies, Graduate School of Frontier Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
Nat Protoc. 2006;1(3):1453-7. doi: 10.1038/nprot.2006.241.
This protocol describes a method for attaching single isolated cardiac myocytes to carbon fibers for mechanical manipulation and measurement. This method relies on cell-adhesive carbon fibers that attach easily to the cell membrane without causing damage, and is thus applicable to intact myocytes. To connect the carbon fiber to micromanipulators, a fiber holder with glass capillaries must first be fabricated. After connection of the fibers to the micromanipulators, firm attachment is easily established by gently pressing the fiber tip onto the cell membrane. Unlike other methods, this technique does not require vast technical expertise, and therefore greatly facilitates experiments. This method enables detection of the effect of drugs, genetic defects or the expression of exogenous proteins on both active and passive properties of cardiac myocytes. In combination with other experimental procedures, this technique can also be applied to the study of mechano-transduction. This protocol can be completed in 3.5 h.
本方案描述了一种将单个分离的心肌细胞附着到碳纤维上以进行机械操作和测量的方法。该方法依赖于易于附着到细胞膜且不会造成损伤的细胞粘附性碳纤维，因此适用于完整的心肌细胞。为了将碳纤维连接到微操纵器上，必须首先制作一个带有玻璃毛细管的纤维固定器。将纤维连接到微操纵器后，通过将纤维尖端轻轻压在细胞膜上即可轻松实现牢固附着。与其他方法不同，该技术不需要大量的技术专业知识，因此极大地促进了实验。该方法能够检测药物、基因缺陷或外源蛋白表达对心肌细胞主动和被动特性的影响。与其他实验程序相结合，该技术还可应用于机械转导的研究。本方案可在3.5小时内完成。
