Name: in vivo 2-photon calcium imaging in layer 2/3 of mice
Uploaded: 2008-03-13T12:00:00
Description: Watch this Scientific Journal Video about In Vivo 2-Photon Calcium Imaging in Layer 2/3 of Mice at JoVE.com

<p>我们要感谢奥尔加Garaschuk有益的讨论，优化大容量装载的钙指标。 </p>

<ol><li>准备的荧光钙指示剂。我们使用acetoxymethyl酯的染料，或AM短染料。这些染料注射细胞外时，可采取由细胞。我们通常使用浓度为1毫米俄勒冈绿BAPTA - 1 AM或荧光凌晨4点。上午染料可从Invitrogen公司在50微克小瓶。首先，我们准备在20％DMSO聚醚F - 127的解决方案。我们热烈的解决方案之前，每天使用，直到其明确的。我们溶解的钙指标的20％为15-20分钟到10毫米的浓度聚醚F-127/DMSO。然后，我们到1mm的解决方案中包含?...

<ol>
	<li>Garaschuk, O., Milos, R. I., Konnerth, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+bulk-loading+of+fluorescent+indicators+for+two-photon+brain+imaging+in+vivo.">Targeted bulk-loading of fluorescent indicators for two-photon brain imaging in vivo.</a> <em style='font-style: italic'>Nat Protoc</em>. <b>1</b>, (2006).</li><li>Stosiek, C., Garaschuk, O., Holthoff, K., Konnerth, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In-vivo+two-photon+calcium+imaging+of+neuronal+networks.">In-vivo two-photon calcium imaging of neuronal networks.</a> <em style='font-style: italic'>Proc Natl Acad Sci USA</em>. <b>100</b>, 7319-7324 (2003).</li><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.&amp;nbsp;&amp;nbsp;+Biomed+Eng+Online+2:13.">ScanImage: flexible software for operating laser scanning microscopes.&amp;nbsp;&amp;nbsp; Biomed Eng Online 2:13.</a> <em style='font-style: italic'>Biomed Eng Online</em>. <b>2</b>, (2003).</li></ol>

<p>电生理记录，这种方法的优势在于它是创伤小，允许在密集的神经元的神经网络活动的录音。执行此过程时，重要的是要记得拿时要格外小心执行了开颅手术，不损伤硬脑膜。即使是少量硬膜下出血，大大模糊的影像。 </p>

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in vivo 2-photon calcium imaging in layer 2/3 of mice

<p class="jove_content">要了解在大脑皮层的微型网络的动态，这是必不可少的，同时记录了大量的神经元活动。在活体双光子钙成像是唯一的方法，允许一个记录密集的人口与单细胞的分辨率神经元活动。该方法包括在植入颅成像窗​​口，可以采取由大量的神经元，最后录制的神经元在体内的双光子显微镜使用时间的推移钙成像活动注入了荧光钙指示剂。星形胶质细胞特异性染料合作注入允许一个有别于星形胶质细胞神经​​元。该技术可以进行小鼠中表达的荧光分子在特定的神经元亚群，以便更好地了解不同的细胞群体的网络交互。 </p>

Watch this Scientific Journal Video about In Vivo 2-Photon Calcium Imaging in Layer 2/3 of Mice at JoVE.com

In Vivo 2-Photon Calcium Imaging in Layer 2/3 of Mice

<p class="jove_content">要了解在大脑皮层的微型网络的动态，这是必不可少的，同时记录了大量的神经元活动。在活体双光子钙成像是唯一的方法，允许一个记录密集的人口与单细胞的分辨率神经元活动。</p>

在体内2光子层钙成像对小鼠的2 / 3

To understand network dynamics of microcircuits in the neocortex, it is essential to simultaneously record the activity of a large number of neurons .  ...

Research

JoVE Journal

Biology

Dissection and 2-Photon Imaging of Peripheral Lymph Nodes in Mice

夹层和小鼠的外周淋巴结的双光子成像

Two-photon imaging has revealed an elegant choreography of motility and cellular interactions within the lymph node under basal conditions and at the ...

科研

生物学

In situ Imaging of the Mouse Thymus Using 2-Photon Microscopy

在胸腺的鼠标采用双光子显微镜的原位成像

Two-photon Microscopy (TPM) enables us to image deep into the thymus and document the events that are important for thymocyte development.  To follow the ...

A Method for 2-Photon Imaging of Blood Flow in the Neocortex through a Cranial Window

通过颅窗口在新皮层的血流量为双光子成像方法

The ability to image the cerebral vasculature (from large vessels to capillaries) and record blood flow dynamics in the intact brain of living rodents is ...

Generation of Bone Marrow Derived Murine Dendritic Cells for Use in 2-photon Imaging

代骨髓衍生的双光子成像使用的小鼠树突状细胞

Several methods for the preparation of murine dendritic cells can be found in the literature. Here, we present a method that produces greater than 85% ...

Imaging Protein-protein Interactions <em>in vivo</em>

Imaging Protein-protein Interactions in vivo

成像蛋白质 - 蛋白质相互作用<em>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, ...

<em>In Vivo</em> 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

<em>在</em>造血干细胞和祖细胞的成年小鼠颅骨骨髓<em>体内</em>四维跟踪

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

<em>En Face</em> Detection of Nitric Oxide and Superoxide in Endothelial Layer of Intact Arteries

En Face Detection of Nitric Oxide and Superoxide in Endothelial Layer of Intact Arteries

一氧化氮和超氧化物歧化酶<em>恩的人脸</em>检测在完整的动脉内皮细胞层

Endothelium-derived nitric oxide (NO) produced from endothelial NO-synthase (eNOS) is one of the most important vasoprotective molecules in cardiovascular ...

Micropuncture of Bowman's Space in Mice Facilitated by 2 Photon Microscopy

微穿刺2光子显微镜辅助小鼠鲍曼空间的研究

Renal micropuncture and renal 2-photon imaging are seminal techniques in renal physiology. However, micropuncture is limited by dependence on conventional ...

Imaging Calcium Dynamics in Subpopulations of Mouse Pancreatic Islet Cells

小鼠胰腺胰岛细胞亚群中的钙动力学成像

Pancreatic islet hormones regulate blood glucose homeostasis. Changes in blood glucose induce oscillations of cytosolic calcium in pancreatic islet cells ...

Identification of Novel Regulators of Plant Transpiration by Large-Scale Thermal Imaging Screening in <em>Helianthus Annuus</em>

Identification of Novel Regulators of Plant Transpiration by Large-Scale Thermal Imaging Screening in Helianthus Annuus

<em>赫连图斯安努斯</em>大型热成像筛选装置新稳压器的鉴定

Plant adaptation to biotic and abiotic stresses is governed by a variety of factors, among which the regulation of stomatal aperture in response to water ...