Ultrashort Pulsed Laser Light for the Manipulation of Neurons and Vasculature

超短激光脉冲作为实验室工具的出现，为探测和操纵生物系统的结构和功能提供了机会。该奖项支持开发一种仪器，该仪器将使用非线性吸收在神经组织中创建微米级的消融，并将附带损害降至最低。该仪器可以与现有的非线性成像设备集成，主要是多光子显微镜。先前的工作表明，超短激光脉冲可以用于组织学的全光学方法，在这种方法中，无需使用机械切片设备即可获得微米级分辨率的解剖结构。试点数据还表明，这种脉冲可以阻止特定的、目标血管中的流动，从而使血管的生物流体力学研究成为可能。所提出的仪器开发计划将设计和实现能够混合光学烧蚀和非线性光学成像的超短脉冲激光技术。这项工作将产生一个近乎交钥匙的系统，该系统使用商业上可获得的激光振荡器和再生放大器，以及商业和定制的光学机械和电子部件和软件的混合。预期的应用包括啮齿动物大脑内神经元和非神经元核团、血管系统和亚细胞结构的自动解剖映射，以及光学分类的诱导，其中切断水蚤神经节中的轴突以研究游泳指挥网络中的电路动力学。其他潜在的应用包括亚微米手术，以消融细胞器，以及短暂破坏血脑屏障。该仪器的设计和实现将使学生能够使用最先进的光学仪器来追求生物学问题。这项拟议的工作将专门培训两名博士后研究人员，学习光学/生物学界面的仪器和实验。

The advent of ultrashort laser light pulses as a laboratory tool has provided the opportunity to probe and manipulate structure and function in biological systems. This award supports development of an instrument that will use nonlinear absorption to create micrometer-sized ablations in nervous tissue, with minimal collateral damage. The instrument can be integrated with existing nonlinear imaging devices, principally the multi-photon microscope. Prior work has shown that ultrashort laser pulses can be used for an all-optical approach to histology in which anatomical structure with micrometer resolution is obtained without the use of mechanical sectioning devices. Pilot data has also demonstrated that such pulses can block flow in specific, targeted blood vessels, thus enabling studies of the biofluid mechanics of the vessels. The proposed instrument development program will design and realize ultrashort pulsed laser technology capable of a mixture of optical ablation and nonlinear optical imaging. The effort will result in a near turn-key system that uses commercially available laser oscillators and regenerative amplifiers, together with a mixture of commercial and custom opto-mechanical and electronic components and software. The expected applications include the automatic anatomical mapping of neuronal and non-neuronal nuclei, vasculature, and subcellular structures within the rodent brain, and the optical induction of axonomy in which axons in the leech ganglion are cut to study circuit dynamics in the swim command network. Additional potential applications include submicrometer surgery to ablate organelles, and transient disruption of the blood-brain barrier. The design and realization of the instrument will expose students to use of state-of the art optical instrumentation for the pursuit of biological questions. The proposed work will specifically train two postdoctoral researchers in instrumentation and experimentation at the optics/biology interface.