Name: in vivo imaging of deep cortical layers using a microprism
Uploaded: 2009-08-27T16:00:00
Description: Watch this Scientific Journal Video about In vivo Imaging of Deep Cortical Layers using a Microprism at JoVE.com

我们感谢安东尼J. Koleske，博士提供YFP的小鼠。

第1部分：微光学显微镜设置<ol><li>我们使用的microprisms 1毫米，直角棱镜，从Optosigma公司（图1）购买的BK7玻璃制成的。 Microprisms处理尽可能使用镊子。 </li><li>斜边的微涂有增强银提供大于97％的反射率从400 - 2000纳米。这提供了内部反射的激光激发光下发出荧光。 </li><li> Microprisms总是在可能时使用产钳处理。手套戴在任何时候都保持棱镜清洁。 </li><li>我们使用一个定制的双光子显微镜的小动物成...

<ol>
	<li>Pologruto, T. A., Sabatini, B. L., 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=ScanImage:+flexible+software+for+operating+laser+scanning+microscopes.">ScanImage: flexible software for operating laser scanning microscopes.</a> Biomed Eng Online. 2, 13-13 (2003).</li><li>Feng, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+neuronal+subsets+in+transgenic+mice+expressing+multiple+spectral+variants+of+GFP.">Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP.</a> Neuron. 28 (1), 41-41 (2000).</li><li>Kleinfeld, D., Mitra, P. P., Helmchen, F., Denk, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluctuations+and+stimulus-induced+changes+in+blood+flow+observed+in+individual+capillaries+in+layers+2+through+4+of+rat+neocortex.">Fluctuations and stimulus-induced changes in blood flow observed in individual capillaries in layers 2 through 4 of rat neocortex.</a> Proc Natl Acad Sci U S A. 95 (26), 15741-15741 (1998).</li></ol>

使用微成像实验相对来说比较简单，在大脑皮层的体内研究提供了许多优点。使用这种技术的有代表性的例子包括在V层皮层神经元的表达黄色荧光蛋白的转基因小鼠拍摄的图像。使用microprisms，人们可以看到一个大V层的锥体细胞机构的收集，近1毫米以下皮质表面。也看到了延长塔夫茨大学（图2）前分歧，通过所有的表面层的顶树突。用于这些实验的老鼠是转基因动物表达YFP的，只有在V层神经元...

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<td>With enhanced silver reflective coating-(R77)</td>
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<td>Fluorescein-dextran</td>
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<td>0.28 NA, 4x air objective</td>
<td>&nbsp;</td>
<td>Olympus</td>
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in vivo imaging of deep cortical layers using a microprism

我们现在活体成像的皮层组织，使用了深刻的脑成像探头在一个微形的协议。 Microprisms 1毫米大小，有一个斜边的反射涂层，让内部的激发和发射光的反射。微探针同时图像多个角度皮质层通常仅见于片的筹备工作。图片是一个大型的收集领域的视图（〜900微米）。此外，我们提供了生存​​的非外科手术的细节和显微镜设置。代表性的结果，包括从你的YFP - H - 1小鼠显示他们的顶树突延长通过表面的皮质层，并延伸到塔夫茨V层锥体神经元的图像。决议足以形象V层神经元的胞体附近的树突棘。一个荧光染料的尾静脉注射，揭示了在皮层血管的复杂网络。流经毛细血管的红细胞（红血球）线扫描揭示RBC速度和通量率可以得到。这种新颖的微探针是一个优雅的，但功能强大的可视化深层细胞的结构和在体内的皮质功能的新方法。

Watch this Scientific Journal Video about In vivo Imaging of Deep Cortical Layers using a Microprism at JoVE.com

In vivo Imaging of Deep Cortical Layers using a Microprism

直角microprisms允许插入到小鼠大脑皮层多个皮质层的深成像与通常在切片中发现了一个观点。一毫米microprisms提供了一个广阔的领域的视图（〜900微米）和空间分辨率足以解决树突棘。我们证明V层神经成像和新皮层的血管成像，使用microprisms。

在使用微深皮质层的活体成像

We present a protocol for in vivo imaging of cortical tissue using a deep-brain imaging probe in the shape of a microprism.  Microprisms are 1-mm in size ...

Research

JoVE Journal

Biology

In vivo Imaging of Deep Cortical Layers using a Microprism

Preparing E18 Cortical Rat Neurons for Compartmentalization in a Microfluidic Device

准备在微流体装置E18条块皮质大鼠神经元

In this video, we demonstrate the preparation of E18 cortical rat neurons. E18 cortical rat neurons are obtained from E18 fetal rat cortex previously ...

科研

生物学

Preparation of Dissociated Mouse Cortical Neuron Cultures

游离的小鼠皮层神经元文化的制备

This video will guide you through the process for generating cortical neuronal cultures from late embryo and early postnatal mouse brain. These cultures ...

Calcium Imaging of Cortical Neurons using Fura-2 AM

使用FURA - 2上午皮层神经元的钙成像

Calcium imaging is a common technique that is useful for measuring calcium signals in cultured cells. Calcium imaging techniques take advantage of calcium ...

In vivo Bioluminescent Imaging of Mammary Tumors Using IVIS Spectrum

In vivo Bioluminescent Imaging of Mammary Tumors Using IVIS Spectrum

使用IVIS频谱的乳腺肿瘤在体内生物发光成像

4T1 mouse mammary tumor cells can be implanted sub-cutaneously in nu/nu mice to form palpable tumors in 15 to 20 days. This xenograft tumor model system ...

Imaging Protein-protein Interactions in vivo

Imaging Protein-protein Interactions in vivo

成像蛋白质 - 蛋白质相互作用在体内</em

Protein-protein interactions are a hallmark of all essential cellular processes. However, many of these interactions are transient, or energetically weak, ...

Single-molecule Imaging of Gene Regulation In vivo Using Cotranslational Activation by Cleavage (CoTrAC)

Single-molecule Imaging of Gene Regulation In vivo Using Cotranslational Activation by Cleavage CoTrAC

单分子成像的基因调控<em在体内</em使用转位激活裂解（CoTrAC）

We describe a fluorescence microscopy method, Co-Translational Activation by Cleavage (CoTrAC) to image the production of protein molecules in live cells ...

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

在造血干细胞和祖细胞的成年小鼠颅骨骨髓体内四维跟踪

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) ...

Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging

蛋白表达和共定位使用复用免疫组织化学染色和多光谱成像定量

Immunohistochemistry is a commonly used clinical and research lab detection technique for investigating protein expression and localization within ...

High-resolution Time-lapse Imaging and Automated Analysis of Microtubule Dynamics in Living Human Umbilical Vein Endothelial Cells

高分辨率成像定时，并在活体人脐静脉内皮细胞微管动力学自动分析

The physiological process by which new vasculature forms from existing vasculature requires specific signaling events that trigger morphological changes ...

Measuring the Stiffness of Ex Vivo Mouse Aortas Using Atomic Force Microscopy

Measuring the Stiffness of Ex Vivo Mouse Aortas Using Atomic Force Microscopy

测量的刚度<em&gt;离体</em&gt;鼠标主动脉用原子力显微镜

Arterial stiffening is a significant risk factor and biomarker for cardiovascular disease and a hallmark of aging. Atomic force microscopy (AFM) is a ...