Name: in vivo two-photon imaging of cortical neurons in neonatal mice
Uploaded: 2018-10-18T14:00:00
Description: Watch this Scientific Journal Video about In Vivo Two-photon Imaging of Cortical Neurons in Neonatal Mice at JoVE.com

作者感谢佐藤、m. 上林和 s. Kouyama 的技术援助。这项工作得到了 jsp KAKENHI 赠款号 JP15K14322 和 JP16H06143、武田科学基金会、上原纪念基金会以及新泻大学脑科学研究所 2017-2923 (陛下) 的合作研究项目的支持, 并由KAKENHI JP16K14559、JP15H01454、JP15H04263 和赠款--在创新领域的科学研究 "从 JP16H06459 (MEXT) 的废料 & 构建系统的动态调节大脑功能" (T.I.)。

实验应按照实验机构规定的动物福利准则进行。
1. 制备用于成像的幼崽
注意: 有稀疏标记的皮层神经元的幼崽可通过超新星载体5,6的子宫电穿孔 (井植敏) 获得。超新星系统由以下两个载体组成: TRE-转基因-转基因-ires-WPRE。在该系统中, 稀疏标记依赖于 TRE 泄漏。在转染神经元的稀疏种群中, TRE 驱动了 Cre 和 ...

<ol>
	<li>Lendvai, B., Stern, E. A., Chen, B., Svoboda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experience-dependent+plasticity+of+dendritic+spines+in+the+developing+rat+barrel+cortex+in+vivo.">Experience-dependent plasticity of dendritic spines in the developing rat barrel cortex in vivo.</a> Nature. 404 (6780), 876-881 (2000).</li><li>Grutzendler, J., Kasthuri, N., Gan, W. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+dendritic+spine+stability+in+the+adult+cortex.">Long-term dendritic spine stability in the adult cortex.</a> Nature. 420 (6917), 812-816 (2002).</li><li>Carrillo, J., Nishiyama, N., Nishiyama, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dendritic+translocation+establishes+the+winner+in+cerebellar+climbing+fiber+synapse+elimination.">Dendritic translocation establishes the winner in cerebellar climbing fiber synapse elimination.</a> The Journal of Neuroscience. 33 (18), 7641-7653 (2013).</li><li>Portera-Cailliau, C., Weimer, R. M., De Paola, V., Caroni, P., Svoboda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diverse+modes+of+axon+elaboration+in+the+developing+neocortex.">Diverse modes of axon elaboration in the developing neocortex.</a> PLoS Biology. , e272 (2005).</li><li>Mizuno, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NMDAR-regulated+dynamics+of+layer+4+neuronal+dendrites+during+thalamocortical+reorganization+in+neonates.">NMDAR-regulated dynamics of layer 4 neuronal dendrites during thalamocortical reorganization in neonates.</a> Neuron. 82 (2), 365-379 (2014).</li><li>Luo, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Supernova:+A+Versatile+Vector+System+for+Single-Cell+Labeling+and+Gene+Function+Studies+in+vivo.">Supernova: A Versatile Vector System for Single-Cell Labeling and Gene Function Studies in vivo.</a> Scientific Reports. 6, 35747 (2016).</li><li>Mizuno, H., Hirano, T., Tagawa, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+for+activity-dependent+cortical+wiring:+formation+of+interhemispheric+connections+in+neonatal+mouse+visual+cortex+requires+projection+neuron+activity.">Evidence for activity-dependent cortical wiring: formation of interhemispheric connections in neonatal mouse visual cortex requires projection neuron activity.</a> The Journal of Neuroscience. 27 (25), 6760-6770 (2007).</li><li>Saito, T., Nakatsuji, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+gene+transfer+into+the+embryonic+mouse+brain+using+in+vivo+electroporation.">Efficient gene transfer into the embryonic mouse brain using in vivo electroporation.</a> Developmental Biology. 240 (1), 237-246 (2001).</li><li>Tabata, H., Nakajima, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+in+utero+gene+transfer+system+to+the+developing+mouse+brain+using+electroporation:+visualization+of+neuronal+migration+in+the+developing+cortex.">Efficient in utero gene transfer system to the developing mouse brain using electroporation: visualization of neuronal migration in the developing cortex.</a> Neuroscience. 103 (4), 865-872 (2001).</li><li>Mizuno, H., Hirano, T., Tagawa, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pre-synaptic+and+post-synaptic+neuronal+activity+supports+the+axon+development+of+callosal+projection+neurons+during+different+post-natal+periods+in+the+mouse+cerebral+cortex.">Pre-synaptic and post-synaptic neuronal activity supports the axon development of callosal projection neurons during different post-natal periods in the mouse cerebral cortex.</a> European Journal of Neuroscience. 31 (3), 410-424 (2010).</li><li>Matsui, A., Yoshida, A. C., Kubota, M., Ogawa, M., Shimogori, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+in+utero+electroporation:+controlled+spatiotemporal+gene+transfection.">Mouse in utero electroporation: controlled spatiotemporal gene transfection.</a> Journal of Visualized Experiments. (54), e3024 (2011).</li><li>Holtmaat, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=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.</a> Nature Protocols. 4 (8), 1128-1144 (2009).</li><li>Nakai, J., Ohkura, M., Imoto, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+high+signal-to-noise+Ca(2+)+probe+composed+of+a+single+green+fluorescent+protein.">A high signal-to-noise Ca(2+) probe composed of a single green fluorescent protein.</a> Nature Biotechnology. 19 (2), 137-141 (2001).</li><li>Chen, T. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasensitive+fluorescent+proteins+for+imaging+neuronal+activity.">Ultrasensitive fluorescent proteins for imaging neuronal activity.</a> Nature. 499 (7458), 295-300 (2013).</li><li>Mizuno, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patchwork-Type+Spontaneous+Activity+in+Neonatal+Barrel+Cortex+Layer+4+Transmitted+via+Thalamocortical+Projections.">Patchwork-Type Spontaneous Activity in Neonatal Barrel Cortex Layer 4 Transmitted via Thalamocortical Projections.</a> Cell Reports. 22 (1), 123-135 (2018).</li><li>Zong, H., Espinosa, J. S., Su, H. H., Muzumdar, M. D., Luo, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mosaic+analysis+with+double+markers+in+mice.">Mosaic analysis with double markers in mice.</a> Cell. 121 (3), 479-492 (2005).</li><li>Young, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single-neuron+labeling+with+inducible+Cre-mediated+knockout+in+transgenic+mice.">Single-neuron labeling with inducible Cre-mediated knockout in transgenic mice.</a> Nature Neuroscience. 11 (6), 721-728 (2008).</li><li>Liu, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neonatal+Repeated+Exposure+to+Isoflurane+not+Sevoflurane+in+Mice+Reversibly+Impaired+Spatial+Cognition+at+Juvenile-Age.">Neonatal Repeated Exposure to Isoflurane not Sevoflurane in Mice Reversibly Impaired Spatial Cognition at Juvenile-Age.</a> Neurochemical Research. 42 (2), 595-605 (2017).</li><li>Kondo, M., Kobayashi, K., Ohkura, M., Nakai, J., Matsuzaki, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two-photon+calcium+imaging+of+the+medial+prefrontal+cortex+and+hippocampus+without+cortical+invasion.">Two-photon calcium imaging of the medial prefrontal cortex and hippocampus without cortical invasion.</a> eLIFE. 6, e26839 (2017).</li><li>Nakazawa, S., Mizuno, H., Iwasato, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+dynamics+of+cortical+neuron+dendritic+trees+revealed+by+long-term+in+vivo+imaging+in+neonates.">Differential dynamics of cortical neuron dendritic trees revealed by long-term in vivo imaging in neonates.</a> Nature Communications. 9 (1), 3106 (2018).</li></ol>

协议和故障排除中的关键步骤:
该协议最关键的步骤是删除头骨 (协议步骤 3.2)。插入时, 刀片通常附着硬脑膜, 导致硬脑膜出血和大脑损伤。这可以避免通过添加一滴皮质缓冲液在头骨和删除头骨在皮质缓冲区。
颅窗准备后, 硬脑膜和皮肤出血导致窗口闭塞。为了避免这种, 在继续下一步之前, 应允许使用组织粘合剂和牙科水泥完全干燥。由于很难完全避免出...

脑皮层神经元电路的精确布线对于更高的大脑功能 (包括知觉、认知、学习和记忆) 至关重要。皮质电路在产后发育过程中动态地被提炼。通过组织学和体外培养分析, 研究了皮层电路的形成过程。然而, 活哺乳动物中的电路形成的动力学仍然主要是未知的。
双光子显微术已广泛应用于成人小鼠脑1、2神经元回路的体内分析。然而, 由于技术上的挑战, 只有有限数量的研究解决了新生小鼠神经元回路的形成。例如, Carrillo et 等执行了在第二个产后周3的小脑中攀爬纤维的延时成像。Portera-他等报道了第一个产后周4皮层层1的轴突成像。在本研究中, 我们总结了一个协议, 观察4层皮层神经元及其在新生小鼠的树突。在我们最近发表的5号出版物中报告了应用本议定书所获得的结果, 其中包括两种方法。首先, 我们使用超新星矢量系统5,6用于标记新生儿大脑中的单个神经元。在超新星系统中, 用于神经元标记的荧光蛋白是可交换的, 标记细胞特异基因敲除和编辑/挖空分析也是可能的。其次, 我们描述了在脆弱的新生小鼠颅窗准备的外科手术。这些方法结合在一起, 允许在体内观察新生儿大脑中的单个神经元。

<table>
	<tbody>
		<tr>
			<td>pK031. TRE-Cre</td>
			<td>Authors</td>
			<td>-</td>
			<td>Available from RIKEN BRC and Addgene</td>
		</tr>
		<tr>
			<td>pK029. CAG-loxP-STOP-loxP-RFP-ires-tTA-WPRE</td>
			<td>Authors</td>
			<td>-</td>
			<td>Available from RIKEN BRC and Addgene</td>
		</tr>
		<tr>
			<td>pK273. CAG-loxP-STOP-loxP-CyRFP-ires-tTA-WPRE</td>
			<td>Authors</td>
			<td>-</td>
			<td>Available from authors</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Wako</td>
			<td>099-06571</td>
			<td></td>
		</tr>
		<tr>
			<td>410 Anaesthesia Unit (isoflurane gas machine)</td>
			<td>Univentor</td>
			<td>8323101</td>
			<td></td>
		</tr>
		<tr>
			<td>Vetbond (tissue adhesive)</td>
			<td>3M</td>
			<td>084-1469SB</td>
			<td></td>
		</tr>
		<tr>
			<td>M&#181;ltiFlex Round (loading tip)</td>
			<td>Sorenson</td>
			<td>13810</td>
			<td></td>
		</tr>
		<tr>
			<td>Gelfoam (gelatin sponge)</td>
			<td>Pfizer</td>
			<td>09-0353-01</td>
			<td></td>
		</tr>
		<tr>
			<td>Agarose</td>
			<td>Sigma</td>
			<td>A9793</td>
			<td>Low melting point</td>
		</tr>
		<tr>
			<td>Round-shaped coverslip</td>
			<td>Matsunami</td>
			<td>-</td>
			<td>Custom made</td>
		</tr>
		<tr>
			<td>Unifast 2 (dental cement)</td>
			<td>GC</td>
			<td>-</td>
			<td></td>
		</tr>
		<tr>
			<td>Titanium bar</td>
			<td>Authors</td>
			<td>-</td>
			<td>Custom made (see Figure 1G)</td>
		</tr>
		<tr>
			<td>Rimadyl (carprofen)</td>
			<td>Zoetis</td>
			<td>-</td>
			<td>Injectable</td>
		</tr>
		<tr>
			<td>2-photon microscope</td>
			<td>Zeiss</td>
			<td>LSM7MP</td>
			<td></td>
		</tr>
		<tr>
			<td>Titanium-sapphire laser</td>
			<td>Spertra-Physics</td>
			<td>Mai-Tai eHPDS</td>
			<td></td>
		</tr>
		<tr>
			<td>Titanium plate</td>
			<td>Authors</td>
			<td>-</td>
			<td>Custom made (see Figure 2A)</td>
		</tr>
		<tr>
			<td>IMARIS, FilamentTracer, MeasurementPro</td>
			<td>BITPLANE</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Goniometer stage</td>
			<td>Thorlabs</td>
			<td>GN2/M</td>
			<td></td>
		</tr>
	</tbody>
</table>

in vivo two-photon imaging of cortical neurons in neonatal mice

双光子成像是一种强大的工具, 用于体内分析哺乳动物大脑中的神经元电路。然而, 存在着有限数量的活体成像方法来检查活的新生哺乳动物的脑组织。本文综述了一种用于活体新生小鼠单个皮层神经元成像的协议。本议定书包括以下两种方法: (1) 在发育中脑皮层神经元稀疏和明亮标记的超新星系统, 以及 (2) 易碎新生儿颅骨的外科手术。该协议允许在高信噪比的新生儿阶段观察单个皮质突起的时间变化。通过将超新星与 RNA 干扰和 CRISPR/Cas9 基因编辑系统相结合, 也可以实现标记细胞特异基因的沉默和挖空。因此, 该协议可用于分析皮层神经元的发展动态、控制神经元动力学的分子机制, 以及疾病模型中神经元动力学的变化。

图 2D - 2F显示了使用本协议的4层皮层神经元双光子时移成像的代表性结果。为了分析, 在整个成像周期中选择具有清晰树突状形态学的神经元。利用形态学分析软件对成像神经元的树突形态学进行了分析。代表性的树突形态学重建如图 2F所示。显示断开连接的树突的神经元 (图 2G) 应排除在分析之外, 因?...

Watch this Scientific Journal Video about In Vivo Two-photon Imaging of Cortical Neurons in Neonatal Mice at JoVE.com

In Vivo Two-photon Imaging of Cortical Neurons in Neonatal Mice

我们提出了一种用于成像新生小鼠大脑皮层的活体双光子成像协议。该方法适用于分析皮层神经元的发育动态、控制神经元动力学的分子机制以及疾病模型神经元动力学的变化。

体内新生小鼠皮层神经元的双光子成像

Two-photon imaging is a powerful tool for the in vivo analysis of neuronal circuits in the mammalian brain. However, a limited number of in vivo imaging ...

这种方法可以帮助回答神经科学中的关键问题，特别是那些与神经元插座形成相关的问题。该技术的主要优点是研究人员可以分析活体新生儿小鼠个体神经元的形态变化。从麻醉后的第五天幼崽开始，只有在对尾部捏紧测试没有反应的情况下才继续。首先，用70%乙醇擦拭头皮，对头皮进行消毒。然后，使用用70%乙醇消毒的剪刀去除覆盖头骨的大约20平方毫米的皮肤。接下来，使用无菌钳子和干净的棉签浸泡在皮层缓冲液中，以去除头骨的筋膜。使用加载尖端将组织粘合剂涂抹在倾斜的皮肤表面，以止血，同时避免要成像的区域。将幼崽放在另一个加热垫上，设置为37摄氏度，并允许它从麻醉中恢复。等待大约 30 分钟，直到组织粘合剂干燥和凝固。一旦组织粘合剂干燥，开始在重新麻醉小鼠的颅窗准备。将一滴皮层缓冲液涂抹在头骨上，然后用无菌剃刀刀片小心地打开头骨直径一毫米的区域，使杜拉完好无损。应用皮层缓冲液以保持大脑表面湿润。使用一小块浸在皮层缓冲液中的明胶海绵来止止出血。使用新鲜片来压缩颅骨切除术的一侧，在不接触杜拉的情况下从硬膜表面排出任何缓冲液和血液。这是准备透明窗口的最关键步骤。杜拉必须完好无损，并且应从暴露的杜拉中去除刷子。接下来，使用移液器尖端涂抹一层薄层百分之一的低熔化红糖，溶解在皮层缓冲液中。将圆形三毫米玻璃盖滑到阿加罗斯凝胶层上。在盖滑和阿加罗斯凝胶层之间浇注过量的阿加罗斯凝胶，以去除所有气泡。使用钳子去除多余的凝胶，从盖滑下突出。混合水泥粉和水泥液体。使用移液器尖端将混合物涂抹在头骨凝固之前。然后，使用牙科水泥将定制的钛棒附在颅骨上。对齐钛条，并平行滑动盖滑，轻松捕获图像。用牙科水泥盖住裸露的头骨。然后，皮下注射镇痛剂。将幼崽放在37摄氏度的加热垫上的恢复笼中一小时，直到牙科水泥凝固。要开始成像，请先设置双光子激光波长。对于 RFP 激发，请使用 1000 纳米的波长。用 70% 乙醇擦拭盖滑的表面。使用钛棒将麻醉幼崽连接到成像阶段的钛板上。使用测角表阶段，调整头部，使盖滑与客观透镜平行。使用设置为 37 摄氏度的加热垫在成像过程中保持幼崽的体温，将异氟兰浓度降低至 0.7% 至 1%将成像阶段置于两光子显微镜的 20 倍客观透镜下。将一滴水涂抹在盖滑上。将软件设置为以 1.4 微米间隔获取 Z 堆栈图像。对于第四层神经元成像，将 Z 宽度设置为 150 到 300 微米，以成像整个树突形态。使用慢速扫描和平均获得显示神经元形态的清晰图像。通常需要超过20分钟才能获得整个树突状形态。此图显示了P5幼崽的四层皮质神经元的代表性Z-堆栈图像。箭头指示要分析的神经元。应从分析中删除树突模糊神经元。此图像显示上一图像中指示神经元的较高放大倍率图像。蓝色箭头指示将在下一个时间点缩回的树突尖，而白色小箭头指示相邻单元格的轴突。4.5 小时后，蓝色箭头的树突尖被收回，带黄色箭头的树突尖被拉长。此处显示了一个具有代表性的树突形态重建，此处显示断开的树突的神经元应排除在分析之外。在尝试此过程时，在过程中保持 dura 完好无损非常重要。使用这个程序，其他方法，如在推进成像可以执行，以回答与新生儿大脑的神经元活动模式相关的其他问题。

Research

JoVE Journal

Neuroscience

In Vivo Two-photon Imaging of Cortical Neurons in Neonatal Mice

A Thin-skull Window Technique for Chronic Two-photon In vivo Imaging of Murine Microglia in Models of Neuroinflammation

A Thin-skull Window Technique for Chronic Two-photon In vivo Imaging of Murine Microglia in Models of Neuroinflammation

一个慢性双光子薄颅骨窗口技术在体内 Neuroinflammation模型小鼠小胶质细胞成像

Traditionally in neuroscience, in vivo two photon imaging of the murine central nervous system has either involved the use of open-skull1,2 or ...

科研

神经科学

In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy

In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy

在体内脊髓的鼠标线使用双光子显微镜成像

In vivo imaging using two-photon microscopy 1 in mice that have been genetically engineered to express fluorescent proteins in specific cell types 2-3 has ...

In Vivo Two-photon Imaging Of Experience-dependent Molecular Changes In Cortical Neurons

In Vivo Two-photon Imaging Of Experience-dependent Molecular Changes In Cortical Neurons

在体内双光子成像的经验依赖的分子在大脑皮质神经元的变化

The brain's ability to change in response to experience is essential for healthy brain function, and abnormalities in this process contribute to a variety ...

In Vivo Two-Photon Microscopy of Single Nerve Endings in Skin

In Vivo Two-Photon Microscopy of Single Nerve Endings in Skin

在皮肤中的单神经末梢体内双光子显微镜

Nerve endings in skin are involved in physiological processes such as sensing1 as well as in pathological processes such as neuropathic pain2. Their ...

Two-Photon in vivo Imaging of Dendritic Spines in the Mouse Cortex Using a Thinned-skull Preparation

Two-Photon in vivo Imaging of Dendritic Spines in the Mouse Cortex Using a Thinned-skull Preparation

双光子<em&gt;在体内</em&gt;树突棘中的鼠标皮质用稀疏的头颅成像准备

In the mammalian cortex, neurons form extremely complicated networks and exchange information at synapses. Changes in synaptic strength, as well as ...

Two-photon Imaging of Cellular Dynamics in the Mouse Spinal Cord

细胞动力学在小鼠脊髓双光子成像

Two-photon (2P) microscopy is utilized to reveal cellular dynamics and interactions deep within living, intact tissues. Here, we present a method for ...

Simultaneous Two-photon In Vivo Imaging of Synaptic Inputs and Postsynaptic Targets in the Mouse Retrosplenial Cortex

Simultaneous Two-photon In Vivo Imaging of Synaptic Inputs and Postsynaptic Targets in the Mouse Retrosplenial Cortex

同时双光子<em&gt;在体内</em&gt;突触输入的影像和突触后目标中的鼠标Retrosplenial的Cortex

This video shows the craniotomy procedure that allows chronic imaging of neurons in the mouse retrosplenial cortex (RSC) using in vivo two-photon ...

Quantification of Filamentous Actin (F-actin) Puncta in Rat Cortical Neurons

Quantification of Filamentous Actin F-actin Puncta in Rat Cortical Neurons

大鼠皮层神经元丝状肌动蛋白（F-肌动蛋白）泪点的量化

Filamentous actin protein (F-actin) plays a major role in spinogenesis, synaptic plasticity, and synaptic stability. Changes in dendritic F-actin rich ...

Two-photon Calcium Imaging in Neuronal Dendrites in Brain Slices

脑切片神经元树突的双光子钙成像

Calcium (Ca2+) imaging is a powerful tool to investigate the spatiotemporal dynamics of intracellular Ca2+ signals in neuronal dendrites. Ca2+ ...

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy

可视化蛋白质激酶 在头部固定行为小鼠中使用 Vivo 双光子荧光寿命成像显微镜的活性

Neuromodulation exerts powerful control over brain function. Dysfunction of neuromodulatory systems results in neurological and psychiatric disorders. ...