Multi-photon nanosurgery in live brain.
Multi-photon nanosurgery in live brain.
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DOI：
10.3389/fnene.2010.00021
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发表时间：
2010-01-01
期刊：
影响因子：
--
通讯作者：
Pavone, Francesco S
中科院分区：
文献类型：
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作者：
Allegra Mascaro, Anna Letizia;Sacconi, Leonardo;Pavone, Francesco S
在过去的几年中，双光子显微镜已被用于在体内进行高空间分辨率成像的神经元，神经胶质细胞和血管结构在完整的新皮层。最近，在平行于其在成像中的应用，多光子吸收已被用作一种工具，在活体动物的神经过程和血管的选择性中断。在这篇综述中，我们提出了一些基本特征的多光子nanosurgery，我们说明了这种新的方法在神经科学研究中提供的优势。我们展示了如何利用多光子激发的空间定位来对表达荧光蛋白的活小鼠的皮层神经元进行选择性损伤。这种方法用于破坏单个神经元，而不会对周围结构造成任何可见的附带损伤。该方法的空间精度允许解剖单个过程以及单个树突棘，保留主要神经元乔木的结构完整性。同样的方法可以通过血管壁的靶向光破坏来突破血脑屏障。我们展示了血管系统如何通过激光消融导致两种不同的中风模型：血管内凝块和外渗。在损伤系统（神经元或血管）通过时间推移体内成像的时间演变之后，可以表征目标结构的生理响应和周围区域的重排。活体脑中的多光子纳米外科手术是产生不同神经退行性疾病模型的有用工具。
In the last few years two-photon microscopy has been used to perform in vivo high spatial resolution imaging of neurons, glial cells and vascular structures in the intact neocortex. Recently, in parallel to its applications in imaging, multi-photon absorption has been used as a tool for the selective disruption of neural processes and blood vessels in living animals. In this review we present some basic features of multi-photon nanosurgery and we illustrate the advantages offered by this novel methodology in neuroscience research. We show how the spatial localization of multi-photon excitation can be exploited to perform selective lesions on cortical neurons in living mice expressing fluorescent proteins. This methodology is applied to disrupt a single neuron without causing any visible collateral damage to the surrounding structures. The spatial precision of this method allows to dissect single processes as well as individual dendritic spines, preserving the structural integrity of the main neuronal arbor. The same approach can be used to breach the blood-brain barrier through a targeted photo-disruption of blood vessels walls. We show how the vascular system can be perturbed through laser ablation leading toward two different models of stroke: intravascular clot and extravasation. Following the temporal evolution of the injured system (either a neuron or a blood vessel) through time lapse in vivo imaging, the physiological response of the target structure and the rearrangement of the surrounding area can be characterized. Multi-photon nanosurgery in live brain represents a useful tool to produce different models of neurodegenerative disease.