Torres-Mapa Maria Leilani, Antkowiak Maciej, Cizmarova Hana, Ferrier David E K, Dholakia Kishan, Gunn-Moore Frank J
Biomed Opt Express. 2011 Jun 1;2(6):1564-75. doi: 10.1364/BOE.2.001564. Epub 2011 May 16.
We demonstrate a system for the combined optical injection and trapping of developing embryos. A Ti:sapphire femtosecond laser in tandem with a spatial light modulator, is used to perform fast and accurate beam-steering and multiplexing. We show successful intracellular delivery of a range of impermeable molecules into individual blastomeres of the annelid Pomatoceros lamarckii embryo by optoinjection, even when the embryo is still enclosed in a chorion. We also demonstrate the ability of the femtosecond laser optoinjection to deliver materials into inner layers of cells in a well-developed embryo. By switching to the continuous wave mode of the Ti:sapphire laser, the same system can be employed to optically trap and orient the 60 μm sized P. lamarckii embryo whilst maintaining its viability. Hence, a complete all-optical manipulation platform is demonstrated paving the way towards single-cell genetic modification and cell lineage mapping in emerging developmental biology model species.
我们展示了一种用于对发育中的胚胎进行光学注射和捕获相结合的系统。一台钛宝石飞秒激光器与一个空间光调制器串联，用于执行快速且精确的光束转向和多路复用。我们通过光注射成功地将一系列不可渗透的分子细胞内递送至环节动物拉氏角贝胚胎的单个卵裂球中，即使胚胎仍包裹在绒毛膜内。我们还证明了飞秒激光光注射能够将材料递送至发育良好的胚胎细胞内层。通过切换到钛宝石激光器的连续波模式，同一系统可用于光学捕获并定向60微米大小的拉氏角贝胚胎，同时保持其活力。因此，展示了一个完整的全光学操作平台，为新兴发育生物学模式物种中的单细胞基因修饰和细胞谱系图谱绘制铺平了道路。
