Kohli Vikram, Elezzabi Abdulhakem Y
9107-116 St, Ultrafast Photonics and Nano-Optics Laboratory, Department of Electrical and Computer Engineering, University of Alberta, Edmonton, T6G2V4, Canada.
BMC Biotechnol. 2008 Jan 29;8:7. doi: 10.1186/1472-6750-8-7.
Femtosecond (fs) laser pulses have recently received wide interest as an alternative tool for manipulating living biological systems. In various model organisms the excision of cellular components and the intracellular delivery of foreign exogenous materials have been reported. However, the effect of the applied fs laser pulses on cell viability and development has yet to be determined. Using the zebrafish (Danio rerio) as our animal model system, we address both the short- and long-term developmental changes following laser surgery on zebrafish embryonic cells.
An exogenous fluorescent probe, fluorescein isothiocyanate (FITC), was successfully introduced into blastomere cells and found to diffuse throughout all developing cells. Using the reported manipulation tool, we addressed whether the applied fs laser pulses induced any short- or long-term developmental effects in embryos reared to 2 and 7 days post-fertilization (dpf). Using light microscopy and scanning electron microscopy we compared key developmental features of laser-manipulated and control samples, including the olfactory pit, dorsal, ventral and pectoral fins, notochord, pectoral fin buds, otic capsule, otic vesicle, neuromast patterning, and kinocilia of the olfactory pit rim and cristae of the lateral wall of the ear.
In our study, no significant differences in hatching rates and developmental morphologies were observed in laser-manipulated samples relative to controls. This tool represents an effective non-destructive technique for potential medical and biological applications.
飞秒（fs）激光脉冲最近作为一种操纵活生物系统的替代工具受到广泛关注。在各种模式生物中，已有报道称可以切除细胞成分并进行外源物质的细胞内递送。然而，所施加的飞秒激光脉冲对细胞活力和发育的影响尚未确定。我们以斑马鱼（Danio rerio）作为动物模型系统，研究了斑马鱼胚胎细胞激光手术后的短期和长期发育变化。
一种外源荧光探针，异硫氰酸荧光素（FITC），成功导入卵裂球细胞，并发现其扩散到所有发育中的细胞中。使用所报道的操纵工具，我们研究了所施加的飞秒激光脉冲是否会在受精后2天和7天（dpf）饲养的胚胎中诱导任何短期或长期发育影响。我们使用光学显微镜和扫描电子显微镜比较了激光操纵样本和对照样本的关键发育特征，包括嗅窝、背鳍、腹鳍和胸鳍、脊索、胸鳍芽、听囊、耳泡、神经丘模式，以及嗅窝边缘的动纤毛和耳侧壁嵴的动纤毛。
在我们的研究中，相对于对照，在激光操纵的样本中未观察到孵化率和发育形态的显著差异。该工具代表了一种有效的非破坏性技术，可用于潜在的医学和生物学应用。
