Okano Kazunori, Wang Chung-Han, Hong Zhen-Yi, Hosokawa Yoichiroh, Liau Ian
Department of Applied Chemistry and Institute of Molecular Science, National Chiao Tung University, Hsinchu 300, Taiwan.
Division of Materials Science, Nara Institute of Science and Technology, Ikoma 630-0192, Japan.
Biochem Biophys Rep. 2020 Oct 6;24:100818. doi: 10.1016/j.bbrep.2020.100818. eCollection 2020 Dec.
The techniques for inducing the death of specific cells in tissue has attracted attention as new methodologies for studying cell function and tissue regeneration. In this study, we show that a sequential process of targeted cell death and removal can be triggered by short-term exposure of near-infrared femtosecond laser pulses. Kinetic analysis of the intracellular accumulation of trypan blue and the assay of caspase activity revealed that femtosecond laser pulses induced immediate disturbance of plasma membrane integrity followed by apoptosis-like cell death. Yet, adjacent cells showed no sign of membrane damage and no increased caspase activity. The laser-exposed cells eventually detached from the substrate after a delay of >54 min while adjacent cells remained intact. On the base of experiments, we applied the same approach to ablate targeted single cardiac cells of a live zebrafish heart. The ability of inducing targeted cell death with femtosecond laser pulses should find broad applications that benefit from precise cellular manipulation at the level of single cells and .
诱导组织中特定细胞死亡的技术作为研究细胞功能和组织再生的新方法已引起关注。在本研究中,我们表明,近红外飞秒激光脉冲的短期照射可触发靶向细胞死亡和清除的连续过程。锥虫蓝细胞内积累的动力学分析和半胱天冬酶活性测定表明,飞秒激光脉冲可立即破坏质膜完整性,随后导致凋亡样细胞死亡。然而,相邻细胞未显示膜损伤迹象,半胱天冬酶活性也未增加。激光照射的细胞最终在延迟超过54分钟后从基质上脱离,而相邻细胞保持完整。基于实验,我们应用相同方法消融活斑马鱼心脏中靶向的单个心肌细胞。飞秒激光脉冲诱导靶向细胞死亡的能力应会有广泛应用,这些应用将受益于单细胞水平的精确细胞操作。