Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window.
Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window.
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DOI：
10.1038/nprot.2009.89
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发表时间：
2009
期刊：
影响因子：
14.8
通讯作者：
Wilbrecht, Linda
中科院分区：
文献类型：
--
作者：
Holtmaat, Anthony;Bonhoeffer, Tobias;Chow, David K.;Chuckowree, Jyoti;De Paola, Vincenzo;Hofer, Sonja B.;Huebener, Mark;Keck, Tara;Knott, Graham;Lee, Wei-Chung A.;Mostany, Ricardo;Mrsic-Flogel, Tom D.;Nedivi, Elly;Portera-Cailliau, Carlos;Svoboda, Karel;Trachtenberg, Joshua T.;Wilbrecht, Linda
为了理解经验依赖性可塑性的细胞和电路机制，需要在完整的大脑中长时间研究神经元及其突触。双光子激发激光扫描显微镜（2 PLSM），连同荧光蛋白的表达，使高分辨率成像的神经元结构在体内。在本协议中，我们描述了一个慢性颅窗，以获得长期成像的小鼠大脑皮层的光学访问。用盖玻片替换小骨瓣，盖玻片用牙科丙烯酸永久密封，提供具有大视野（0.8-12 mm 2）的清晰成像窗口。手术过程可在101小时内完成。该制备允许以任意成像间隔在数月的时间段内成像。大尺寸的成像窗口有利于小鼠中小神经元结构的持续结构可塑性的成像，具有低密度的标记神经元。单个神经元的整个树突和轴突乔木可以重建。
To understand the cellular and circuit mechanisms of experience-dependent plasticity, neurons and their synapses need to be studied in the intact brain over extended periods of time. Two-photon excitation laser scanning microscopy (2PLSM), together with expression of fluorescent proteins, enables high-resolution imaging of neuronal structure in vivo. In this protocol we describe a chronic cranial window to obtain optical access to the mouse cerebral cortex for long-term imaging. A small bone flap is replaced with a coverglass, which is permanently sealed in place with dental acrylic, providing a clear imaging window with a large field of view (∼0.8–12 mm2). The surgical procedure can be completed within ∼1 h. The preparation allows imaging over time periods of months with arbitrary imaging intervals. The large size of the imaging window facilitates imaging of ongoing structural plasticity of small neuronal structures in mice, with low densities of labeled neurons. The entire dendritic and axonal arbor of individual neurons can be reconstructed.
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DOI：
10.1073/pnas.0810149105
发表时间：
2008-12-16
影响因子：
11.1
作者：
Lee, Wei-Chung Allen;Chen, Jerry L.;Nedivi, Elly
通讯作者：
Nedivi, Elly
影响因子：
25
作者：
Chow, David K.;Groszer, Matthias;Pribadi, Mochtar;Machniki, Michal;Carmichael, S. Thomas;Liu, Xin;Trachtenberg, Joshua T.
通讯作者：
Trachtenberg, Joshua T.
影响因子：
16.2
作者：
Holtmaat, AJGD;Trachtenberg, JT;Svoboda, K
通讯作者：
Svoboda, K
影响因子：
64.8
作者：
Lendvai, B;Stern, EA;Svoboda, K
通讯作者：
Svoboda, K
影响因子：
9.8
作者：
Lee WC;Huang H;Feng G;Sanes JR;Brown EN;So PT;Nedivi E
通讯作者：
Nedivi E