König K, Riemann I, Fischer P, Halbhuber K J
Institute of Anatomy II, Friedrich Schiller University Jena, Germany.
Cell Mol Biol (Noisy-le-grand). 1999 Mar;45(2):195-201.
We report on laser-assisted knocking out of genomic nanometer-sized regions within the nucleus of living cells. The intranuclear nanosurgery was possible by application of highly intense near infrared femtosecond laser pulses. The non-contact laser treatment was performed within a closed sterile cell chamber. The destructive multiphoton effect was based on 10(12) W/cm2 light intensities and limited to a sub-femtoliter focal volume of a high numerical aperture objective. We used the intracellular nanoscalpel for highly precise non-contact dissection of Hoechst-labelled chromosomes within a nucleus of a living Chinese hamster ovary cell. Following laser treatment, the cell remained alive and did not show any signs of membrane perturbation. The use of near infrared pulses provide the possibility of non-invasive intracellular nanoprocessing also within living tissue in depths of more than 100 microns.
我们报告了在活细胞核内利用激光辅助敲除基因组纳米级区域的研究。通过应用高强度近红外飞秒激光脉冲，实现了核内纳米手术。非接触式激光治疗在封闭的无菌细胞室内进行。破坏性多光子效应基于10¹²W/cm²的光强，且局限于高数值孔径物镜的亚飞升焦体积内。我们使用细胞内纳米手术刀对活的中国仓鼠卵巢细胞核内经Hoechst标记的染色体进行高精度非接触切割。激光处理后，细胞仍存活，且未表现出任何膜扰动迹象。使用近红外脉冲还为在深度超过100微米的活组织内进行非侵入性细胞内纳米加工提供了可能性。
