Name: in vivo functional brain imaging approach based on bioluminescent calcium indicator gfp-aequorin
Uploaded: 2016-01-08T15:00:01
Description: Watch this Scientific Journal Video about In Vivo Functional Brain Imaging Approach Based on Bioluminescent Calcium Indicator GFP-aequorin at JoVE.com

We are indebted to E. Karplus, from Sciences Wares, USA, for his precious and useful help and advices. We thank E. Carbognin, A. Avet-Rochex, M. Murmu, P. Pavot, D. Minocci, G. Vinatier, and J. Stinnakre for their contribution to the development and improvement of this technique. We also thank B. Pfeiffer and G. Rubin, Janelia Farm, H.H.M.I., Ashburn, USA, for the pJFRC65-13XLexAop2-IVS-G5A-BP line. This work was supported by the Chateaubriand Fellowship, French Embassy, Washington, USA to A. Lark, by the National Science Foundation (IOS 1352882) to T. Kitamoto, and by the French ANRs (ANR-05-NEUR-009 Drosaequorin, 2005, and FlyBrainImaging, 2011), the Conseil R&#233;gional Ile-De France (NeRF and DIM), the IFR-144, the Physique-Chimie-Biologie Interface Program of the CNRS (2009), and by the CNRS, France to JR Martin.

1.样品的制备<ol><li>溶液的制备和设置<ol><li>保持在标准的食品中，在24℃的所有果蝇线。后与他们保持低密度小瓶生成标准化的尺寸和重量的苍蝇。 <ol><li>添加10处女女性，10名男性在小瓶中，让它们交配，每2天他们转移到一个新的小瓶。然后用10天后，当苍蝇开始E关闭，每天收获苍蝇。保持苍蝇的年龄的准确记录，并让他们在良好的状态。 </li><li>在第3天，分开女性的男性和保持女性20?...

<ol>
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T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+biogenic+amine+and+a+neuropeptide+act+identically:+tyramine+signals+through+calcium+in+Drosophila+tubule+stellate+cells.">A biogenic amine and a neuropeptide act identically: tyramine signals through calcium in Drosophila tubule stellate cells.</a> Proc. Biol. Sci. 280 (1757), 20122943 (2013).</li><li>Leclerc, C., Webb, S. E., Daguzan, C., Moreau, M., Miller, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+patterns+of+calcium+transients+during+neural+induction+in+Xenopus+laevis+embryos.">Imaging patterns of calcium transients during neural induction in Xenopus laevis embryos.</a> J. Cell Sci. 113 (19), 3519-3529 (2000).</li><li>Martin, J. -. R., Rogers, K. L., Chagneau, C., Br&#251;let, P. <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+bioluminescence+imaging+of+Ca2++signalling+in+the+brain+of+Drosophila.">In vivo bioluminescence imaging of Ca2+ signalling in the brain of Drosophila.</a> PLoS One. 2 (3), e275 (2007).</li><li>Murmu, M. S., Stinnakre, J., Martin, J. -. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Presynaptic+Ca2++stores+contribute+to+odor-induced+responses+in+Drosophila+olfactory+receptor+neurons.">Presynaptic Ca2+ stores contribute to odor-induced responses in Drosophila olfactory receptor neurons.</a> J. Exp. Biol. 213 (24), 4163-4173 (2010).</li><li>Murmu, M. S., Stinnakre, J., R&#233;al, E., Martin, J. -. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calcium-stores+mediate+adaptation+in+axon+terminals+of+Olfactory+Receptor+Neurons+in+Drosophila.">Calcium-stores mediate adaptation in axon terminals of Olfactory Receptor Neurons in Drosophila.</a> BMC Neurosci. 12, 105 (2011).</li><li>Pavot, P., Carbognin, E., Martin, J. -. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PKA+and+cAMP/CNG+channels+independently+regulate+the+cholinergic+Ca2+-response+of+Drosophila+mushroom+body+neurons.">PKA and cAMP/CNG channels independently regulate the cholinergic Ca2+-response of Drosophila mushroom body neurons.</a> eNeuro. 2 (2), 0054-0068 (2015).</li><li>Minocci, D., Carbognin, E., Murmu, M. S., Martin, J. -. R. <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+functional+calcium+imaging+of+induced+or+spontaneous+activity+in+the+fly+brain+using+a+GFP-apoaequorin-based+bioluminescent+approach.">In vivo functional calcium imaging of induced or spontaneous activity in the fly brain using a GFP-apoaequorin-based bioluminescent approach.</a> BBA-Mol Cell Res. 1833 (7), 1632-1640 (2013).</li><li>Lima, S. Q., Miesenb&#246;ck, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Remote+control+of+behavior+through+genetically+targeted+photostimulation+of+neurons.">Remote control of behavior through genetically targeted photostimulation of neurons.</a> Cell. 121 (1), 141-152 (2005).</li><li>Shimomura, O., Musicki, B., Kishi, Y., Inouye, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Light-emitting+properties+of+recombinant+semisynthetic+aequorins+and+recombinant+fluorescein-conjugated+aequorin+for+measuring+cellular+calcium.">Light-emitting properties of recombinant semisynthetic aequorins and recombinant fluorescein-conjugated aequorin for measuring cellular calcium.</a> Cell Calcium. 14 (5), 373-378 (1993).</li><li>Berridge, M. J., Bootman, M. D., Roderick, H. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calcium+signalling:+dynamics,+homeostasis+and+remodelling.">Calcium signalling: dynamics, homeostasis and remodelling.</a> Nat Rev Mol Cell Bio. 4 (7), 517-529 (2003).</li><li>Fiala, A., Spall, T. <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+calcium+imaging+of+brain+activity+in+Drosophila+by+transgenic+cameleon+expression.">In vivo calcium imaging of brain activity in Drosophila by transgenic cameleon expression.</a> Sci STKE. 2003 (174), PL6 (2003).</li><li>Wang, J. W., Wong, A. M., Flores, J., Vosshall, L. B., Axel, R. <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+reveals+an+odor-evoked+map+of+activity+in+the+fly+brain.">Two-photon calcium imaging reveals an odor-evoked map of activity in the fly brain.</a> Cell. 112 (2), 271-282 (2003).</li><li>Wilson, R. I., Turner, G. C., Laurent, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transformation+of+Olfactory+Representations+in+the+Drosophila+Antennal+Lobe.">Transformation of Olfactory Representations in the Drosophila Antennal Lobe.</a> Science. 303 (5656), 366-370 (2004).</li></ol>

这里介绍的最近开发的生物发光型绿色荧光蛋白水母发光蛋白的方法允许在 钙 -activity在不同的神经元体内功能记录功能，如果需要的话，以及在其他类型的细胞，如肾星状细胞的报道中卡夫雷罗等。 2013年5。 修改后，故障排除，附加部件及关键步骤 果蝇畜牧 <p class="jove_cont...

基本构图和大脑及其离散结构中的活性作用长期以来一直是神经科学领域内的紧张的学习领域。也许有人用来解决这个问题的最早的成功做法是在电活动变化的直接生理测量 - 但是替代方法，允许电压，pH或钙离子浓度的变化（作为活动代理）的实时成像带来了额外的功能，研究神经元活动1。成像技术携带的优点各种各样如降低侵袭和全脑结构内监控活性的能力。此外，在动物听话遗传学，包括果蝇 ，遗传编码的钙指标，如卡默莱昂，GCaMPs和其他发展1已使研究人员能够监测单个神经元，神经元群体和全脑结构。而更传统的荧光钙成像方法使用GCaMPs（GFP融合至钙调蛋白）或FRET（CFP和YFP融合至钙调蛋白）已被证明是提供良好的空间和时间分辨率有用的技术，光激发的基础过程滋生于它的实验一些限制应用程序。由于光激发，自体荧光，光漂白，并且光毒性的性质都不可避免地产生，这进而限制了可以被成像的结构的深度，记录的持续时间，以及记录的实时连续性。换句话说，记录必须经常进行间歇地避免这些不希望的副作用。 因为这些挑战的，​​已经开发了钙成像的其他替换的方法。其中最有希望的方法之一是生物发光成像，这依赖于通过后的钙结合的酶反应产生的光。 Bioluminescen牛逼成像不需要光激发，因此不会遇到同样的困难，传统的钙成像。所述生物发光性报告水母发光蛋白-从水母中分离，同样水母从中GFP也isolated-经由它的三个的EF手结构结合钙和发生构象变化，从而导致其辅因子腔肠素的氧化和蓝光的释放（λ = 469纳米）2,3。水母发光蛋白具有钙（K D =10μM），这使得它适合用于监测的钙瞬变，因为它产生非常少的背景噪声和不与细胞内钙离子的缓冲系统 4干扰非常低的亲和力。虽然水母发光蛋白以前已用于监测神经诱导的过程在爪蟾卵5产生的光太暗用于神经元活动的实时成像。为了努力应对这一挑战，菲利普登记及＃251;让我们和他的同事在巴斯德研究所创建的基因构建融合GFP和水母发光蛋白基因，水母4中模拟天然状态。此融合基因产物经由水母发光蛋白的三个的EF手结构，其经历构象变化，导致了腔肠素，其中以非辐射到GFP传送能量的氧化再次结合钙。这种能量转移的结果，而不是蓝色的光从水母发光蛋白从GFP绿光（λ= 509纳米）的释放。 GFP和水母发光蛋白的融合也赋予更高的蛋白质稳定性帮助融合蛋白产生光比单独4水母发光蛋白明亮的19至65倍。使用大脑内生物发光方法扩展了无论是在连续记录的持续时间和结构的可记录大脑内的数表示在当前的实验应用钙成像的。概括地说在以前的工作中就意味着全球性和更大的各种大脑的区域化“活性”可以被监视（通过钙瞬变的代理）。更重要的是活性可为更长的被监测，在实时和连续的基础上，这很容易适合于追求基底函数的大脑中的完整的理解。 在过去的几年里，我们的实验室和其他人已开发转基因果蝇表达二进制表达系统的控制下的绿色荧光蛋白水母发光蛋白（GAL4 / UAS-GFP-水母发光蛋白）5,6。我们使用GFP的水母发光蛋白的生物发光通过自然刺激如香味7,8，以及调制的 Ca 2+ -activity在以下乙酰胆碱受体的刺激通过直接烟碱应用9蘑菇体研究了细胞成分产生的记录活动。此外，我们还使用GFP的水母发光蛋白，以解决了一些一直无法被预先处理实验问题，如长期成像自发钙瞬变和更深的大脑结构10的可视化。最近，我们已经使用了GAL4 / UAS系统表达的哺乳动物的ATP受体P2X 2 11的阳离子通道，在投射神经元（PNS）和使用的LexA的系统表达的GFP-水母发光蛋白在蘑菇体（MB247-LexA的， 13XLexAop2-IVS-G5A-BP）（礼节性B.法伊弗，珍妮莉娅法姆，美国）。这使我们通过应用ATP，从而激活蘑菇体（期票的下游靶标），以激活期票，模仿更自然的条件下，如响应一个刺激气味。总体来说，这一技术结合P2X 2和GFP-水母扩大实验的可能性。从广义上它提供了机会，以更好地理解活动模式在大脑中，开始对不同的刺激和干扰如何改变活动的基础图案的新研究。

<table>
	<tbody>
		<tr>
			<td>NaCl</td>
			<td>Sigma-Aldrich</td>
			<td>S3014</td>
			<td></td>
		</tr>
		<tr>
			<td>CaCl2</td>
			<td>Merck</td>
			<td>1023911000</td>
			<td></td>
		</tr>
		<tr>
			<td>HEPES</td>
			<td>Sigma-Aldrich</td>
			<td>7365-45-9</td>
			<td></td>
		</tr>
		<tr>
			<td>KCL&#160;</td>
			<td>prolabo</td>
			<td>26 764.298</td>
			<td>normapur</td>
		</tr>
		<tr>
			<td>MgCl</td>
			<td>prolabo</td>
			<td>25 108.295</td>
			<td>normapur</td>
		</tr>
		<tr>
			<td>sucrose</td>
			<td>Sigma-Aldrich</td>
			<td>S0389</td>
			<td></td>
		</tr>
		<tr>
			<td>Nicotine</td>
			<td>Sigma-Aldrich</td>
			<td>N3876</td>
			<td></td>
		</tr>
		<tr>
			<td>ATP</td>
			<td>Sigma-Aldrich</td>
			<td>A2383</td>
			<td>Adenosine 5&#8242;-triphosphate disodium salt hydrate</td>
		</tr>
		<tr>
			<td>benzyl-coelenterazine</td>
			<td>Prolume, nanolight&#160;</td>
			<td>Cat #301 Coelenterazine h</td>
			<td>http://www.prolume.com/</td>
		</tr>
		<tr>
			<td>dental glue</td>
			<td>3M ESPE&#8482;</td>
			<td>46954</td>
			<td>Protemp&#8482; IV</td>
		</tr>
		<tr>
			<td>silicon glue</td>
			<td>3M ESPE&#8482;</td>
			<td>36958</td>
			<td>Express&#8482; 2 Regular Body Quick</td>
		</tr>
		<tr>
			<td>tape</td>
			<td>3M Scotch</td>
			<td>122s</td>
			<td>3M Scotch Magic Tape</td>
		</tr>
		<tr>
			<td>pipette tips</td>
			<td>Corning Incorporated</td>
			<td>4868</td>
			<td>100-1000&#181;L Univesal Pipette Tip</td>
		</tr>
		<tr>
			<td>chamber and holder (stage)</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>hand made</td>
		</tr>
		<tr>
			<td>incubation box</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>hand made</td>
		</tr>
		<tr>
			<td>forceps (No 5)</td>
			<td>Fine Science Tools</td>
			<td>11252-00</td>
			<td>Micro-dissecting forceps</td>
		</tr>
		<tr>
			<td>curved forceps</td>
			<td>Sigma-Aldrich</td>
			<td>F4142</td>
			<td>Micro-dissecting forceps</td>
		</tr>
		<tr>
			<td>glue applicator&#160;</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>hand made&#160;</td>
		</tr>
		<tr>
			<td>knife</td>
			<td>Fine Science Tools</td>
			<td>10316-14</td>
			<td>5mm Depth 15&#176; stab</td>
		</tr>
		<tr>
			<td>EM-CCD camera&#160;</td>
			<td>Andor</td>
			<td>DU-897E-CS0-#BV</td>
			<td>iXon (cooled to -80&#176;C)</td>
		</tr>
		<tr>
			<td>Microscope&#160;</td>
			<td>Nikon</td>
			<td>N/A</td>
			<td>Eclipse-E800</td>
		</tr>
		<tr>
			<td>immersion objective lens</td>
			<td>Nikon</td>
			<td>N/A</td>
			<td>20x Fluor .5w</td>
		</tr>
		<tr>
			<td>dissection microscope with fluorescence</td>
			<td>Leica MZ Fl III</td>
			<td>N/A</td>
			<td>leica dissection scope and florescent lamp</td>
		</tr>
		<tr>
			<td>tight dark box</td>
			<td>Science Wares</td>
			<td>N/A</td>
			<td>Custom built by Science Wares</td>
		</tr>
		<tr>
			<td>peristaltic pump&#160;</td>
			<td>Gilson</td>
			<td>N/A</td>
			<td>Minipuls 2</td>
		</tr>
		<tr>
			<td>tubing</td>
			<td>Fisher Scientific</td>
			<td>depends on thickness</td>
			<td>Tygon R3603, St-Gobain</td>
		</tr>
		<tr>
			<td>perfusion system</td>
			<td>Warner</td>
			<td>64-0135 &#160;(VC-66CS)</td>
			<td>Perfusion valve control system, complete, with pinch valves, 6 channel</td>
		</tr>
		<tr>
			<td>perfusion regulators</td>
			<td>Leventon</td>
			<td>201108</td>
			<td>Dosi-flow 3</td>
		</tr>
		<tr>
			<td>measurement and automation explorer</td>
			<td>Sciences Wares &#38; National Instruments</td>
			<td>N/A</td>
			<td>software http://sine.ni.com/nips/cds/view/p/lang/en/nid/1380</td>
		</tr>
		<tr>
			<td>photon imager&#160;</td>
			<td>Science Wares &#38; National Instruments</td>
			<td>N/A</td>
			<td>software http://sine.ni.com/nips/cds/view/p/lang/en/nid/1380</td>
		</tr>
		<tr>
			<td>photon viewer</td>
			<td>Science Wares &#38; National Instuments</td>
			<td>N/A</td>
			<td>software http://sine.ni.com/nips/cds/view/p/lang/en/nid/1380</td>
		</tr>
		<tr>
			<td>Excel</td>
			<td>Microsoft</td>
			<td>N/A</td>
			<td>software https://www.microsoft.com</td>
		</tr>
		<tr>
			<td>virtual dub</td>
			<td>open access&#160;</td>
			<td>N/A</td>
			<td>software http://virtualdub.sourceforge.net/</td>
		</tr>
		<tr>
			<td>UAS-GA2</td>
			<td>Martin et al., 2007, Ref. 1&#160;</td>
			<td>N/A</td>
			<td>&#160;No stocks {currently} publicly available&#160;</td>
		</tr>
		<tr>
			<td>pJFRC65-13XLexAop2-IVS-G5A-BP&#160;&#160;</td>
			<td>B. Pfeiffer, Janelia Farms, Ashburn USA.&#160;</td>
			<td>(STOCK #1117340)</td>
			<td>pfeifferb@janelia.hhmi.org</td>
		</tr>
		<tr>
			<td>P2X2</td>
			<td>Lima and Misenboeck, Ref. 11&#160;</td>
			<td>N/A</td>
			<td>&#160;No stocks {currently} publicly available&#160;</td>
		</tr>
		<tr>
			<td>OK107</td>
			<td>Bloomington stock center</td>
			<td>854</td>
			<td>http://flystocks.bio.indiana.edu/</td>
		</tr>
	</tbody>
</table>

in vivo functional brain imaging approach based on bioluminescent calcium indicator gfp-aequorin

功能体内成像已经成为一个强大的方法来研究功能和脑细胞和利益结构的生理机能。近来已经开发的 Ca 2+使用生物发光性报告的GFP-水母发光蛋白（GA）-imaging的一种新方法。这种新的技术依赖于的绿色荧光蛋白和水母发光蛋白的基因融合，产生分子能够结合钙和 - 与另外的辅助因子腔肠中 - 发光明亮的光线可以通过光子收集器进行监控。已经产生对小鼠和果蝇携带GFP的水母发光蛋白基因的转基因品系，在 果蝇中，绿色荧光蛋白水母发光蛋白基因已被放置在GAL4 / UAS二进制表达系统允许大脑内针对性表达和成像的控制之下。这个方法后来被证明是能够检测两个向内的 Ca 2+ -transients和Ca 2+从内存储-released。最重要的是它允许在连续记录，成像在大脑内的更深的深度更大的持续时间，并记录在高时间分辨率（高达8.3毫秒）。在这里，我们提出了利用生物发光成像记录和分析钙蘑菇体中2+ -activity，结构中心在飞大脑学习和记忆的基本方法。

将GFP对水母发光蛋白的融合可以让我们通过GFP的激发形象化我们之前生物发光成像的兴趣区域在显微镜上（图3A，4A，6A，6E）荧光模式。一个刺激的蘑菇体的最简单的方法是通过激活离子型乙酰胆碱受体。虽然乙酰胆碱是该受体的内源性配体，我们已发现，尼古丁产生在蘑菇体更具有可重复性的响应。这部分是因为它们特异性表达的烟碱乙酰胆碱受体，从而通过使用尼古?...

Watch this Scientific Journal Video about In Vivo Functional Brain Imaging Approach Based on Bioluminescent Calcium Indicator GFP-aequorin at JoVE.com

In Vivo Functional Brain Imaging Approach Based on Bioluminescent Calcium Indicator GFP-aequorin

在基于生物发光钙指标GFP的水母体内脑功能成像方法

在这里，我们提出了一个新颖的钙利用生物发光记者-imaging方法。这种方法使用的稠构建GFP-水母发光蛋白，其结合的 Ca 2+并发光，省去了光激发。显著此方法允许长时间连续成像，获得脑深部的结构和高时间分辨率。

Functional in vivo imaging has become a powerful approach to study the function and physiology of brain cells and structures of interest. Recently a new ...

Research

JoVE Journal

Neuroscience

In Vivo Functional Brain Imaging Approach Based on Bioluminescent Calcium Indicator GFP-aequorin

Functional Calcium Imaging in Developing Cortical Networks

在开发皮层网络功能钙成像

A hallmark pattern of activity in developing nervous systems is spontaneous, synchronized network activity. Synchronized activity has been observed in ...

科研

神经科学

Functional Neuroimaging Using Ultrasonic Blood-brain Barrier Disruption and Manganese-enhanced MRI

使用超声波血脑屏障破坏和锰增强MRI的脑功能成像

Although mice are the dominant model system for studying the genetic and molecular underpinnings of neuroscience, functional neuroimaging in mice remains ...

In vivo Neuronal Calcium Imaging in C. elegans

In vivo Neuronal Calcium Imaging in C. elegans

C.在体内神经元钙成像技术线虫

The nematode worm C. elegans is an ideal model organism  for relatively simple, low cost neuronal imaging in vivo. Its small transparent body and simple, ...

Simultaneous Electrophysiological Recording and Calcium Imaging of Suprachiasmatic Nucleus Neurons

Simultaneous electrophysiological and fluorescent imaging recording methods were used to study the role of changes of membrane potential or current in ...

Ex Vivo Calcium Imaging for Visualizing Brain Responses to Endocrine Signaling in Drosophila

Ex Vivo Calcium Imaging for Visualizing Brain Responses to Endocrine Signaling in Drosophila

前体钙成像用于可视化大脑对果蝇内分泌信号的反应

Organ-to-organ communication by endocrine signaling, for example, from the periphery to the brain, is essential for maintaining homeostasis. As a model ...

In Vivo Calcium Imaging of Lateral-line Hair Cells in Larval Zebrafish

幼虫斑马鱼外系头发细胞的活体钙成像

Sensory hair cells are mechanoreceptors found in the inner ear that are required for hearing and balance. Hair cells are activated in response to sensory ...

In vivo Calcium Imaging in Mouse Inferior Olive

In vivo Calcium Imaging in Mouse Inferior Olive

在体内 小鼠劣质橄榄中的钙成像

Inferior olive (IO), a nucleus in the ventral medulla, is the only source of climbing fibers that form one of the two input pathways entering the ...

&#181;Tongue: A Microfluidics-Based Functional Imaging Platform for the Tongue In Vivo

Ãƒâ€šÃ‚ÂµTongue: A Microfluidics-Based Functional Imaging Platform for the Tongue In Vivo

μTongue：一个基于微流体的功能成像平台，用于在Viva的舌头

Intravital fluorescence microscopy is a tool used widely to study multicellular dynamics in a live animal. However, it has not been successfully used in ...

Brain Pericyte Calcium and Hemodynamic Imaging in Transgenic Mice In Vivo

Brain Pericyte Calcium and Hemodynamic Imaging in Transgenic Mice In Vivo

活体转基因小鼠脑围产层钙和血液学成像

Recent advances in protein biology and mouse genetics have made it possible to measure intracellular calcium fluctuations of brain cells in vivo and to ...

Stereotaxic Viral Injection and Gradient-Index Lens Implantation for Deep Brain In Vivo Calcium Imaging

Stereotaxic Viral Injection and Gradient-Index Lens Implantation for Deep Brain In Vivo Calcium Imaging

A miniature fluorescence microscope (miniscope) is a potent tool for in vivo calcium imaging from freely behaving animals. It offers several advantages ...