Patterned femtosecond-laser ablation of Xenopus laevis melanocytes for studies of cell migration, wound repair, and developmental processes.
Patterned femtosecond-laser ablation of Xenopus laevis melanocytes for studies of cell migration, wound repair, and developmental processes.
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DOI：
10.1364/boe.2.002383
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发表时间：
2011-08-01
影响因子：
3.4
通讯作者：
Omenetto FG
中科院分区：
文献类型：
--
作者：
Mondia JP;Adams DS;Orendorff RD;Levin M;Omenetto FG
超快（飞秒）激光已经成为研究生物现象的重要工具，因为它们能够在外科应用中实现高度局部的组织去除。在这里，我们描述了在非洲爪蟾蝌蚪上发现的可编程、微尺度、飞秒激光消融黑素细胞，这项技术适用于发育、再生和癌症研究的生物学研究。我们说明了单个黑素细胞的激光标记，并在黑素细胞簇上绘制图案，以帮助跟踪它们的迁移和/或再生。我们还证明，该系统可以通过清除黑素细胞区域并监测其随时间的恢复，将刮伤测试（一种广泛用于培养细胞研究细胞迁移和伤口愈合的技术）升级到更现实的体内领域。此外，我们展示了黑素细胞消融如何通过损伤邻近组织来用于功能丧失实验，以局部脊髓损伤后尾巴再生异常为例。由于黑素细胞的大小、形状和深度随蝌蚪年龄和黑素细胞位置（头部或尾部）的变化而变化，因此给出了消融阈值图。讨论了激光烧蚀的机理。
Ultrafast (femtosecond) lasers have become an important tool to investigate biological phenomena because of their ability to effect highly localized tissue removal in surgical applications. Here we describe programmable, microscale, femtosecond-laser ablation of melanocytes found on Xenopus laevis tadpoles, a technique that is applicable to biological studies in development, regeneration, and cancer research. We illustrate laser marking of individual melanocytes, and the drawing of patterns on melanocyte clusters to help track their migration and/or regeneration. We also demonstrate that this system can upgrade scratch tests, a technique used widely with cultured cells to study cell migration and wound healing, to the more realistic in vivo realm, by clearing a region of melanocytes and monitoring their return over time. In addition, we show how melanocyte ablation can be used for loss-of-function experiments by damaging neighboring tissue, using the example of abnormal tail regeneration following localized spinal cord damage. Since the size, shape, and depth of melanocytes vary as a function of tadpole age and melanocyte location (head or tail), an ablation threshold chart is given. Mechanisms of laser ablation are also discussed.