Name: real-time bioluminescence imaging of notch signaling dynamics during murine neurogenesis
Uploaded: 2019-12-12T13:00:00
Description: Watch this Scientific Journal Video about Real-time Bioluminescence Imaging of Notch Signaling Dynamics during Murine Neurogenesis at JoVE.com

我们感谢岩本裕子支持制作该视频。我们也感谢伊索拉伊郎对图像分析的讨论和支持，宫崎骏为转基因动物的生成提供技术支持，Yuji Shinjo（奥林匹斯医学科学），江川正雄（奥林匹斯医学科学），石津（石津）奥林巴斯医学）和吴昆田基（日本安多）为生物发光成像系统的技术支持和讨论。这项工作得到了进化科学和技术核心研究（JPMJCR12W2）（R.K.）、创新领域科学研究资助（H.S.MEXT 24116705和R.K.MEXT 16H06480）的支持，科学研究资助（C）的支持。18K06254（H.S.），日本文部科学省，以及日本教育、文化、体育、科学和技术部的动态生活系统方法平台。

所有程序，包括动物科目，都通过了京都大学前沿生命与医学研究所的动物护理和使用委员会的批准。
1. 生物发光记者
注： 荧光素酶报通过融合降解信号，可用于测量启动子活性的快速动态。此外，荧光素酶融合报告器能够监测单个细胞中的蛋白质动力学。这两种类型的报告可用于单层培养（分离培养）和组织培养（切片培养）实验。
<ol><li>记者监测...

<ol>
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Notch信号的分量在发生过程中显示振荡表达同步，但在神经发生期间不同步，导致在后一种情况下通过静态分析捕获表达动力学的困难。因此，需要实时监控来揭示 Notch 信令分量（如Hes1和Dll1）的表达动态。由于Hes1和Dll1振荡的表达周期极短，因此需要大约 2-3 小时、快速响应和不稳定的报告器来监测其表达动态。为此，我们开发了生物发光报告仪和成像系统。生物发光?...

哺乳动物的大脑由大量不同类型的神经元和胶质细胞组成。所有细胞都是从神经干细胞/祖细胞（NPC）生成的，这些细胞首先增殖以扩大其数量，然后开始分化成神经元，最后产生胶质细胞1、2、3、4、5。一旦细胞分化成神经元，它们就不能增殖或增加其数量，因此，在后期阶段维持NPC是很重要的。通过细胞-细胞相互作用的槽口信令在维持NPC6，7中起着重要的作用。凹槽配体与邻近细胞表面的膜蛋白Notch相互作用，并激活Notch蛋白。激活后，诺奇蛋白发生蛋白解解，从而将Notch（NICD）的细胞内域从细胞膜释放到细胞核8、9、10中。在核中，NICD与Hes1和Hes5（Hes1/5）的启动子区域结合，并激活这些基因的表达。 Hes1/5抑制前神经基因Ascl1和Neurogenin1/2（Neurog1/2）的表达11，12，13，14。由于前神经基因诱导神经元分化，Hes1/5在维持NPC中起着至关重要的作用。此外，由于前神经基因可以激活诺奇配体三角洲（Dll1）的表达，Hes1/5还抑制Dll1的表达。因此，Dll1的表达导致通过Notch信令对Dll1的相邻细胞呈阴性。这样，细胞会抑制相邻细胞遵循其相同的命运，这种现象称为横向抑制8。在发育中的大脑中，横向抑制在产生各种不同的细胞类型方面起着作用。
在单个单元水平上的实时成像揭示了NPC15、16、17中Notch信号分量的动态表达。Notch信号激活Hes1的表达，但Hes1蛋白与它自己的启动子结合并抑制它自己的表达。此外，Hes1是一种极不稳定的蛋白质，由泛基蛋白-蛋白酶体通路降解;因此，对自身促进者的压制只是短暂的，然后转录又开始了。这样，Hes1的表达在2小时周期18的转录和平移水平上振荡。Hes1的振荡表达，反过来，通过周期性抑制15，16，17，19，诱导下游目标基因的振荡表达，如Ascl1，Neurog2和Dll1。 虽然前神经基因可以诱导神经元分化，但振荡表达不足以进行神经元分化;而是它们的持续表达对神经元分化至关重要。前神经基因的振荡表达对于维持NPC很重要，而不是诱导神经元分化14，15，16。Dll1的表达在各种形态形成（如神经发生和躯体发生）期间在转录和转化水平上振荡。Dll1的动态表达对Dll1的正常形态发生和稳定表达具有重要的诱导神经发生缺陷和躯体发生缺陷17。这些发现证明了基因表达和蛋白质动力学对调节各种发育事件（即不同的表达动力学在细胞行为中产生不同输出）的重要作用。
为了分析Notch信号的动态，组织和细胞的静态分析是不够的，因为它们在不断变化。单细胞的实时成像是揭示基因表达动态的有力工具。Notch信号分子的动态表达在2-3小时周期内发生快速循环反应。这种快速的周期性表达为实时监测带来了两个难题：（1）分子的表达被抑制到低水平，（2）快速周转需要快速响应的记者从。为了克服这些问题，我们之前开发了一种生物发光实时成像方法20。由于生物发光报告仪比荧光报仪具有更高的灵敏度和较短的成熟时间，因此这种策略使我们能够监测活细胞的快速动力学。使用实时可视化，我们发现更多的基因表现出动态表达比我们之前认为的。此外，显示活细胞表达和蛋白质动力学的报告数量以及这些动力学在各种生物事件中的重要性有所增加，这表明动力学在基因表达中的基本作用21，22。
在本报告中，我们描述了一种在分离区域性和皮质切片培养中可视化 NPC 中 Notch 配体 Dll1 表达式的方法。为了监测单细胞水平Dll1转录的动态，我们产生了从携带pDll1-Ub-Fluc报告器的转基因小鼠的胚胎端管中衍生的NPC分离培养物，该实验鼠是Dll1启动子驱动的不稳定的荧光酶报告者。为了监测体内的Dll1蛋白动力学，我们将Dll1-Fluc融合报告器引入皮层中的NPC，并可视化了报告器在皮质切片培养中的NPC的表达。实时成像使我们能够以高时间分辨率捕捉活细胞中基因表达和蛋白质动力学的各种特征。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Bioluminescence Imaging System</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Chilled water circulator (chiller)</td>
			<td>Julabo</td>
			<td>Model: F12-ED</td>
			<td></td>
		</tr>
		<tr>
			<td>Cooled CCD camera</td>
			<td>Andor Technology</td>
			<td>Model: iKon-M 934</td>
			<td></td>
		</tr>
		<tr>
			<td>Incubator system</td>
			<td>TOKAI HIT</td>
			<td>Model: INU-ONICS</td>
			<td></td>
		</tr>
		<tr>
			<td>Inverted microscope</td>
			<td>Olympus</td>
			<td>Model: IX81</td>
			<td></td>
		</tr>
		<tr>
			<td>Inverted microscope</td>
			<td>Olympus</td>
			<td>Model: IX83</td>
			<td></td>
		</tr>
		<tr>
			<td>LED illumination device</td>
			<td>CoolLED</td>
			<td>Model: pE1</td>
			<td></td>
		</tr>
		<tr>
			<td>MetaMorph</td>
			<td>MOLECULAR DEVICES</td>
			<td>Model: 40000</td>
			<td></td>
		</tr>
		<tr>
			<td>Mix gas controller</td>
			<td>Tokken</td>
			<td>Model: TK-MIGM OLO2</td>
			<td></td>
		</tr>
		<tr>
			<td>Objective lens</td>
			<td>Olympus</td>
			<td>Model: UPLFLN 40X O</td>
			<td></td>
		</tr>
		<tr>
			<td>Preparations for Dissection</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dissection microscope</td>
			<td>Nikon</td>
			<td>Model: SMZ-2B</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluorescence stereoscopic microscope</td>
			<td>Leica</td>
			<td>Model: MZ16FA</td>
			<td></td>
		</tr>
		<tr>
			<td>Fine forceps</td>
			<td>DUMONT</td>
			<td>INOX No.5</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors, Micro scissors</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ring-shaped forceps</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>10-cm plastic petri dish</td>
			<td>greiner</td>
			<td>664160-013</td>
			<td></td>
		</tr>
		<tr>
			<td>35-mm plastic petri dish</td>
			<td>greiner</td>
			<td>627160</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>Nacalai Tesque</td>
			<td>14249-24</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM/F12</td>
			<td>invitrogen</td>
			<td>11039-021</td>
			<td></td>
		</tr>
		<tr>
			<td>Reagents for NPC dissociation culture</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>B27 supplement</td>
			<td>invitrogen</td>
			<td>12587-010</td>
			<td></td>
		</tr>
		<tr>
			<td>bFGF</td>
			<td>invitrogen</td>
			<td>13256-029</td>
			<td>Stock solution: 1 &#956;g/ml in 0.1% BSA/PBS</td>
		</tr>
		<tr>
			<td>D-luciferin</td>
			<td>Nacalai Tesque</td>
			<td>01493-85</td>
			<td>Stock solution: 100mM in 0.9% saline</td>
		</tr>
		<tr>
			<td>DNase</td>
			<td>Worthington Biochemical Corporation</td>
			<td>LK003172</td>
			<td>Stock solution: 1000U/ml in EBSS</td>
		</tr>
		<tr>
			<td>EBSS</td>
			<td>Worthington Biochemical Corporation</td>
			<td>LK003188</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass bottom dish</td>
			<td>IWAKI</td>
			<td>3910-035</td>
			<td></td>
		</tr>
		<tr>
			<td>N2 supplement (100x)</td>
			<td>invitrogen</td>
			<td>17502-048</td>
			<td></td>
		</tr>
		<tr>
			<td>N-acetyl-cystein</td>
			<td>Sigma</td>
			<td>A-9165-25G</td>
			<td></td>
		</tr>
		<tr>
			<td>Papain</td>
			<td>Worthington Biochemical Corporation</td>
			<td>LK003178</td>
			<td>Stock solution: 7U/ml in EBSS</td>
		</tr>
		<tr>
			<td>Penicillin/Streptmycine</td>
			<td>Nacalai Tesque</td>
			<td>09367-34</td>
			<td></td>
		</tr>
		<tr>
			<td>Poly-L-lysine</td>
			<td>Sigma</td>
			<td>P-6281</td>
			<td>40 mg/ml in DW</td>
		</tr>
		<tr>
			<td>Preparations for in utero electroporation</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>50-ml syringe</td>
			<td>TERUMO</td>
			<td>181228T</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrode</td>
			<td>Neppagene</td>
			<td>7-mm</td>
			<td></td>
		</tr>
		<tr>
			<td>Electroporator</td>
			<td>Neppagene</td>
			<td>CUY21 EDIT</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Gauzes</td>
			<td>Kawamoto co.</td>
			<td>7161</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro capillary</td>
			<td>Made in-house</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>Nacalai Tesque</td>
			<td>14249-24</td>
			<td></td>
		</tr>
		<tr>
			<td>Pentbarbital</td>
			<td>Kyoritsuseiyaku</td>
			<td>Somnopentyl</td>
			<td></td>
		</tr>
		<tr>
			<td>Ring-shaped forceps</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors, Micro scissors</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Suture needle</td>
			<td>Akiyama MEDICAL MFG. CO</td>
			<td>F17-40B2</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Bayer</td>
			<td>Seractal</td>
			<td></td>
		</tr>
		<tr>
			<td>Preparations for Slice culture</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>10-cm plastic petri dish</td>
			<td>greiner</td>
			<td>664160-013</td>
			<td></td>
		</tr>
		<tr>
			<td>35-mm plastic petri dish</td>
			<td>greiner</td>
			<td>627160</td>
			<td></td>
		</tr>
		<tr>
			<td>Culture insert</td>
			<td>Millipore</td>
			<td>PICM01250</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM/F12</td>
			<td>invitrogen</td>
			<td>11039-021</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum</td>
			<td>Sigma</td>
			<td>172012-500ML</td>
			<td></td>
		</tr>
		<tr>
			<td>Fine forceps</td>
			<td>DUMONT</td>
			<td>INOX No.5</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Horse Serum</td>
			<td>Gibco</td>
			<td>16050-122</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro surgical knife</td>
			<td>Alcon</td>
			<td>19 Gauge V-Lance</td>
			<td></td>
		</tr>
		<tr>
			<td>Multi-gas incubator</td>
			<td>Panasonic</td>
			<td>MCO-5MUV-PJ</td>
			<td></td>
		</tr>
		<tr>
			<td>N2/B27 media</td>
			<td>Made in-house</td>
			<td></td>
			<td>ref. NPC dissociatioin culture</td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>Nacalai Tesque</td>
			<td>14249-24</td>
			<td></td>
		</tr>
		<tr>
			<td>Ring-shaped forceps</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors, Micro scissors</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Silicon rubber cutting board</td>
			<td>Made in-house</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

real-time bioluminescence imaging of notch signaling dynamics during murine neurogenesis

Notch信号通过细胞-细胞相互作用调节神经干细胞/祖细胞的维持。Notch 信令的组件表现出动态表达式。槽口信号效应器 Hes1 和 Notch 配体增量类似 1 （Dll1） 在神经茎/祖细胞中以振荡方式表示。由于这些基因的振荡表达周期很短（2小时），因此很难监测它们的循环表达。为了检查基因表达或蛋白质动力学的这种快速变化，需要快速响应报告器。由于其快速成熟动力学和高灵敏度，生物发光报明酶适用于监测活细胞中快速的基因表达变化。我们使用不稳定的荧光素酶报告器来监测促进者活动，使用荧光素酶融合报告器在单个细胞分辨率下可视化蛋白质动力学。这些生物发光记者显示快速周转，并产生非常微弱的信号;因此，我们开发了一个高度敏感的生物发光成像系统来检测这种微弱的信号。这些方法使我们能够监测活细胞和组织的各种基因表达动力学，这是帮助了解实际细胞状态的重要信息。

基因Hes1/7的表达表现出2小时振荡周期，在各种细胞系和细胞生成过程中。此外，振荡周期很短，其mRNA和蛋白质都极不稳定，半寿命在20分钟左右。如果使用反应缓慢的报告器，我们就无法追踪这种快速的动态，如果使用稳定的报告器，它会在基因表达振荡时逐渐积累。因此，记者必须迅速退化，以监测这些周期性表达基因的快速流动。为了克服这些问题，我们使用荧光酶报告器来监测振荡...

Watch this Scientific Journal Video about Real-time Bioluminescence Imaging of Notch Signaling Dynamics during Murine Neurogenesis at JoVE.com

Real-time Bioluminescence Imaging of Notch Signaling Dynamics during Murine Neurogenesis

神经干细胞/祖细胞表现出Notch信号分量的各种表达动力学，导致细胞事件的不同结果。这种动态表达可以通过实时监测，而不是静态分析来显示，使用高度敏感的生物发光成像系统，能够可视化基因表达的快速变化。

毛因神经发生期间诺奇信号动力学的实时生物发光成像

Notch signaling regulates the maintenance of neural stem/progenitor cells by cell-cell interactions. The components of Notch signaling exhibit dynamic ...

该方法有助于回答有关基因表达动力学在细胞增殖和分化中的重要性，特别是神经干细胞中诺奇信号分子动力学的关键问题。这项技术的主要优点是，我们可以通过活体成像以非常高的精度监测体外和体内的基因表达。演示这个程序的将是我实验室的助理教授HiromiShimojo，他开发了这项新技术。对于Dll1 F荧光素记者介绍进入NPC，确认在胚胎日12.5至14.5怀孕小鼠的踏板反射缺乏反应，然后通过腹部，沿中线进行2至3厘米切口。将 PBS 浸泡的纱布放在切口周围，并使用环状钳子小心地提取右子宫角。数出胚胎后，使用微毛细管将一至两微升混合Dll1 F荧光石分析器DNA注入每个胚胎的心灵脑室。成功的注射将导致在子宫表面遵守蓝色染料。对于电穿孔，用PBS将子宫和电极弄湿，轻轻抓住第一个胚胎的头部。将带正电荷的电极设置为注入DNA的半球一侧，并提供5个30至50伏、50毫秒的脉冲，脉冲之间暂停一秒，检查带负电荷的电极是否产生气泡。当所有胚胎都注射后，将左子宫角返回到腹部，并使用17毫米的针头用4-0丝缝合线关闭切口，在完成闭合前将温暖的PBS放入腹腔。为了建立分离的NPC培养，首先将子宫从胚胎12.5天到14.5日怀孕的Dll1泛小通通荧光素转基因小鼠放入含有25毫升冰冷PBS的10厘米培养皿中，并使用微剪刀和细钳将胚胎从子宫中取出。斩首胚胎后，将头部放入含有冰冷 DMEM/F12 的新的 10 厘米培养皿中，并去除每个大脑周围的表皮和软骨。在含有三毫升冷N2B27介质的解剖显微镜下将大脑转移到35毫米的培养皿中。从死头，从右和左电脑中去除。使用细钳去除覆盖电脑表面的脑毛，并从电脑中解剖皮层的侧侧部分。当所有感兴趣的神经组织被分离后，使用 P1000 移液器将样品转移到单独的 1.5 毫升管中，并使用 P200 移液器从每个管中吸出任何额外的介质。每皮层添加100微升的木瓜溶液，并将样品在24摄氏度下放置15分钟。在孵化结束时，用新的P1000移液器轻轻移液样品10次，将管子返回24摄氏度，再增加15分钟。在孵育结束时，再次轻轻移液样品，然后通过离心收集消化的组织。吸制超自然剂，用一毫升DMEM/F12介质轻轻重新暂停每个颗粒。将样品离心三次，每次离心后，用新鲜毫升的介质轻轻重新补充每个颗粒。上次离心后，将N2B27介质的500微升颗粒重新加用1毫摩尔荧光素，将每个样品中的6个细胞种子种子到单独的聚L-Lysine涂层玻璃底盘上。然后，将板放在细胞培养培养箱中一小时。一旦细胞粘附在盘子上，在每盘中加入两毫升新鲜N2B27培养基加一毫升荧光素。要可视化在NPC分离文化中的荧光酶记者表达，选择油浸物目标，将样品盘放在显微镜舞台上。手动查看该字段以确定查看单元格的最佳位置，并专注于感兴趣的单元格。单击"实时"获取测试图像，并使用多维采集程序通过二维发光和亮场采集运行延时采集 24 小时。为了准备发展皮层切片培养，记者注射后一天，采集胚胎13.5至14.5胚胎大脑，将含有大脑的菜放在荧光立体显微镜的舞台上。确认每个皮层在适当的激发光下表达注射的荧光报告器，然后将大脑转移到一个硅橡胶切割板上，该切割板充满30毫升的DMEM/F12介质，气泡为100%氧气。使用微型手术刀和细钳切割后脑和横向部分之间的边界，将组织分离成两个半球。然后，使用刀切割皮层像条纹，以获得皮质切片，转移切片在中等从切割板到一个新的35毫米培养皿，因为他们被获取。收集所有切片后，将每个切片的切割表面放在包含富集的玻璃底盘上，并相应地用细钳调整每个切片的方向。然后，用移液器去除多余的介质，将培养皿放入设置为 40% 氧气、5%二氧化碳和 37 摄氏度的多气体培养箱中，30 分钟。在孵化结束时，在培养皿中加入300个富营养介质微升，并辅以一毫摩尔荧光素。要可视化切片培养物中的荧光酶处理器表达式，请选择 40 倍目标，然后将样品盘放在显微镜舞台上。获取荧光的测试图像，并在适当的激发光的照明下将位置和焦点平面设置为感兴趣的区域。然后，通过三维发光、荧光和亮场采集运行延时采集 24 小时。为了监测振荡基因Dll1的启动者活动，可以使用泛素荧光酶，一个半寿命约10分钟的不稳定荧光素酶记者。像荧光报告器一样，荧光酶报告器可以通过与感兴趣的基因编码序列融合来监测蛋白质的表达动力学。为了在组织培养中单细胞水平可视化记者表达，如所证明，记者基因可以通过子宫电穿孔短暂转染到NPC中。此外，基于显微镜的成像可实现对亮场、荧光和化学发光图像的采集。Dll1推广活动记者展示了NPC中源自Dll1泛化F百诗记者小鼠的电脑的振荡表达，不稳定的荧光酶记者表示启动者活动表达的上下剧烈调节。在相邻的增强绿色荧光蛋白阳性细胞中，携带 Hes1 记者和 mCherry 阳性细胞表示 Dll1 蛋白质报告器在观察过程中彼此接触，Hes1 记者的表达似乎在接触后大约 60 分钟开始，表明相邻细胞之间传递 Notch 信号的时间延迟大约一个小时。此外，在信号传输过程中，Dll1蛋白质表达表现出动态转移。获得生物发光成像时，保持显微镜室完全暗，因为无关的光线可以防止最佳获得。光遗传学的最新发展使我们能够用光来控制基因表达。通过该技术，我们可以同时引入各种基因表达模式，并监测荧光素酶报告器的反应。

Education

Research

JoVE Core

JoVE Journal

Cell Biology

Molecular Biology

Developmental Biology

Unit 2

Unit 1

Cell Signaling Pathways

Alternative Signaling Routes in Gene Expression

SCIENTISTS IN ACTION

Studying Wnt Signaling During Patterning of Conducting Airways

研究Wnt信号传导过程中的航空公司图案

Wnt signaling pathways play critical roles during development of the respiratory tract. Defining precise mechanisms of differentiation and morphogenesis ...

科研

发育生物学

Kinetic Measurement and Real Time Visualization of Somatic Reprogramming

体细胞重编程的动力学测量和实时可视化

Somatic reprogramming has enabled the conversion of adult cells to induced pluripotent stem cells (iPSC) from diverse genetic backgrounds and disease ...

Stimulation of Notch Signaling in Mouse Osteoclast Precursors

在鼠标破骨细胞前体Notch信号的刺激

Notch signaling is a key component of multiple physiological and pathological processes. The nature of Notch signaling, however, makes in vitro ...

Multi-Photon Time Lapse Imaging to Visualize Development in Real-time: Visualization of Migrating Neural Crest Cells in Zebrafish Embryos

多光子时间推移成像实时可视化发展： 迁移神经嵴细胞在斑马鱼胚胎中的可视化

Congenital eye and craniofacial anomalies reflect disruptions in the neural crest, a transient population of migratory stem cells that give rise to ...

Real-time Measurement of Epithelial Barrier Permeability in Human Intestinal Organoids

人肠道 Organoids 上皮屏障通透性的实时测量

Advances in 3D culture of intestinal tissues obtained through biopsy or generated from pluripotent stem cells via directed differentiation, have resulted ...

In Vivo Imaging of Muscle-tendon Morphogenesis in Drosophila Pupae

In Vivo Imaging of Muscle-tendon Morphogenesis in Drosophila Pupae

在体内果蝇蛹肌肌腱形态的影像学研究

Muscles together with tendons and the skeleton enable animals including humans to move their body parts. Muscle morphogenesis is highly conserved from ...

Live Cell Imaging of Chromosome Segregation During Mitosis

有丝分裂过程中染色体分离的活体细胞成像

Chromosomes must be reliably and uniformly segregated into daughter cells during mitotic cell division. Fidelity of chromosomal segregation is controlled ...

Neurogenesis Using P19 Embryonal Carcinoma Cells

使用P19胚胎癌细胞的神经发生

The P19 cell line derived from a mouse embryo-derived teratocarcinoma has the ability to differentiate into the three germ layers. In the presence of ...

Imaging and Analysis of Tissue Orientation and Growth Dynamics in the Developing Drosophila Epithelia During Pupal Stages

Imaging and Analysis of Tissue Orientation and Growth Dynamics in the Developing Drosophila Epithelia During Pupal Stages

普帕尔阶段发育性皮瘤中组织定向和生长动力学的成像与分析

Within multicellular organisms, mature tissues and organs display high degrees of order in the spatial arrangements of their constituent cells. A ...

Real-Time Imaging of CCL5-Induced Migration of Periosteal Skeletal Stem Cells in Mice

CCL5诱导小鼠骨膜骨骼干细胞迁移的实时成像

Periosteal skeletal stem cells (P-SSCs) are essential for lifelong bone maintenance and repair, making them an ideal focus for the development of ...