笔者想感谢米歇尔·德奇为完善这一眼消融技术，泰勒·伯克霍尔茨与功能测试援助，迈克尔·勒文的抗抑制蛋白抗体，而顺治Morokuma为珀尔帖板信息。这项工作是由西密歇根大学WSB的SFSA资助。

1.动物文化和处理注:此协议使用Schmidtea地中海 ，二倍体涡物种，基因组测序16，17是常用的再生研究。然而，该测定法是与其他物种，诸如Girardia虎纹和Girardia dorotocephala（它们是可商购）同样成功。 <ol><li> 保持"蠕虫水"为0.5g / L海盐超纯（或过滤的去离子）水制成蠕虫。使用无菌聚丙烯或玻璃容器来保存蠕虫水。见<a href="http://ecsource.jove.com/files/ftp_upload/55594/Supplementary_File_1.docx" target="_blank">补充文件1</a>对蠕虫水制备的细节。 注意:不要使用肥皂清洗容器（或其它电源）如肥皂是有毒的蠕虫;代替用70％乙醇擦拭，并允许风干。 &lt;OL&gt; <li>戴上手套，用涡虫的工作，以保护他们免受污染的同时。 注:蠕虫是环境毒素和化学品，包括肥皂，漂白剂，洗发水，护发素，和洗手液非常敏感。 </li></ol></li><li>房子菌落在聚丙烯食品容器，与盖子部分打开在约20℃（室温）下的空气交换。在任一培养皿商店实验蠕虫（每100mm培养皿20个蠕虫）或未经处理的组织培养板（每孔1个蠕虫，对于12孔板）。为了减轻压力，保持双方的殖民地和实验在黑暗中。 注:殖民地应该有每升水虫不超过500-1000蠕虫。对于实验，让完全到位盖子，由于气流是专为这些菜。 </li><li>通过从移液管滴加果泥，放置食物的液滴在整个容器订阅与泥有机牛肉或鸡肉肝脏非实验蠕虫每周一次。允许蠕虫2个小时以消耗食物在清理容器在天黑前（参见步骤1.4）。在使用之前，储存在-20℃下短期（周）原浆或-80℃长期（数月）;不重新冻结泥再利用（如细菌可以开花损害蠕虫）。 注意事项:肝应该不含激素或抗生素，最好不预先冻结。冷冻前（前馈或）离心机原浆以除去空气并防止食物浮动。饮食宜沉到容器的底部。原浆的1毫升足以用于500-1,000蠕虫。 </li><li>兑水每周一次用新鲜水虫两个殖民地和实验。菌落的容器也应与未漂白纸巾擦拭（以除去粘液残基陷阱废物）每周一次（供给和再次2天后后2小时）饥饿蠕虫，或每周两次用于馈送蠕虫。为了留住生物膜，不抹容器的80％以上。 </li><li> 移动容器（或手术SETU之间蠕虫p）的使用移液管。到附着于容器表面自由蠕虫，喷出的水过/蜗杆以从表面释放它的后面。然后吸取蜗杆到移液管，而试图保持在移液管的底部英寸（较薄）部分内的蜗杆。 <ol><li>后送吸管吮吸起来之前，有少量虫水。 </li><li>尝试移动水绕的蠕虫病毒，而不是病毒本身，以帮助防止用吸管触及他们撕裂软体蠕虫。 </li><li>保持覆盖，除非转让蠕虫或更换水实验菜肴。 </li></ol></li></ol> 2.准备工作<ol><li> 停止至少一周之前的实验喂养蚯蚓。 <ol><li>选择已没喂食了≥1周并且至少5-7毫米长度蠕虫。转移蠕虫至培养皿2/3满蠕虫水的，并通过滑动盘下方的标尺确认虫长度。测量w ^奥姆斯同时充分延伸和移动。 注:虽然小（5-7毫米）蠕虫执行后续免疫荧光原位杂交分析，更大的蠕虫（8-10毫米）时，更容易与工作更好地工作-尤其是当学习消融。 </li><li>确保蠕虫是整体，无破损或最近再生。 注意:作为相对于正常蠕虫蠕虫再生将在头部和/或尾部区轻得多（或没有）颜料。 </li><li>如果蠕虫进行RNAi或药物治疗，在这一点上做到这一点。如果蠕虫被馈送作为处理（作为RNA干扰）的一部分，等待最后喂食后至少7天继续到步骤2.2之前。 </li></ol></li><li> 清洁用70％乙醇的工作空间和解剖显微镜基，并允许它完全干燥。选择蠕虫的盘放置到所述范围的左边。到的范围的权利，放置一对＃5钳子的，一个31克5 / 16-英寸胰岛素针用1毫升的注射器，和一个干净的移液管。 <ol><li>确保器械接触到工作台（其可以引入污染物）保持。使用硬质物品（如筷子休息），以保持镊子，针和吸管干净。可替换地，在新鲜棕色（未漂白的）纸巾的顶部位置的仪器。 注:因为它们与右手这些个人和随后的指令写入。左手个人应该扭转左/右方向。 </li></ol></li><li>旁边的工作空间，将无绒织物擦巾的盒子，新鲜水虫的来源，以及一些额外的移液管（从台式保护）。地方虫水的塑料洗瓶便于分配的。还放置一个标记的12孔板（或100毫米培养皿）的范围的权利，用于收集被消融的动物。 </li><li> 如果使用定制的珀耳帖板来固定蠕虫，珀耳帖板定位到在叔凹陷他解剖显微镜的基部和直到珀耳帖板的工作表面是充分地冷却调整直流电源上的输出（通常为〜5 V）。可选地，通过填充100毫米培养皿使冷板½充分用水和冷冻至少24小时。丢弃该盖板，将解剖范围的基础上的100mm培养皿的底部，然后将60毫米的培养皿的盖颠倒直接在冰（图1A）的表面上。 <ol><li>水在100mm培养皿已经冻结后，冰的"火山"可以出现在板的中心。如果发生这种情况，用一个刀片刮冰平，以从表面除去任何不规则性。 注:在手术的冰会融化，使得消融困难得多。预先准备几个冰盘，并更换冻结的用于尽快60毫米培养皿的盖开始浮动在测定过程中熔化的。 </li></ol></li><li> 准备手术的表面。切5厘米×10厘米的塑料片石蜡膜（4cm 2的如使用培养皿冷板）并将其放置在固定装置的中心。折叠无绒布纸巾擦拭成方形大致2 cm 2并且将其放置在膜的顶部。切下一块的白色滤纸1.5-2厘米2，将其上的擦拭顶部。参见图1B-1E。 <ol><li>按住折叠处用戴手套的手指擦拭，并用虫水轻轻挫伤擦拭，以确保它保持折叠。使用移液管的一侧滚动抹平。 注意:冷的温度有助于保持蠕虫移动。工作"干"也有助于固定。组织擦拭行为通过滤纸芯吸水，手术过程中保持蜗杆湿润而不使蠕虫完全风干（这是致命的）。 </li></ol></li></ol> 3.手术消融<ol><li> 用移液管上放置È蠕虫背侧上到滤纸上。在关闭光源和引导鹅颈，使光线集中在蠕虫。调整解剖范围聚焦和放大倍率使眼睛清晰可见（整个蠕虫并不需要在图）。放大5倍是一个很好的起点。通过使得头部指向朝向研究员（朝向显微镜的前面）倾斜30-40度的石蜡膜旋转，向右定向蠕虫。 <ol><li>避免使用明亮的光线，以防止多余的蠕虫运动。涡虫显示负趋光性，并会尝试逃避耀眼的光芒。 </li><li>如果蠕虫被定位朝上（含咽口可见）腹侧，使用产钳无光泽的一面轻轻地重新定位蠕虫背侧了（眼睛可见）。避免接近动物用钳子的尖端，以尽量减少重新定位动物时通过软组织撕裂的可能性。 </li><li&gt;调整显微镜焦点使眼睛显然聚焦。此外，还要确保两个眼睛和周围的乳头组织在视野中。 </li></ol></li><li> 保持新鲜针/注射器在右手就好像它是一个笔（拇指和食指之间）。做好防珀尔帖板（或培养皿冷板）的左拇指，并使用左拇指为支点在其上的注射器的底部将休息（图2A）。通过显微镜观察，并确保针的斜面是可见的。 注:针是非常尖锐的。操作时他们一定要小心。戴乳胶或丁腈手套可以提供一些保护。 <ol><li>使用左手拇指，以帮助保持针/注射器在整个手术过程平稳，避免握手。 </li><li>成大约40°用左手拇指和左手食指（与手术表面上的食指）的角度，以帮助保持手术的表面稳定。 </li></ol></li><li> 同朝上（如调羹）所述针的斜面，定位在垂直于眼（图2B）的针。使用针的尖端轻轻穿透组织覆盖在眼睛的视杯（白色，未着色区域）的薄层，由右至左舀。非常温和地去除位于视杯内的任何色素斑组织，小心不要刺破底部或破坏视杯边界。 <ol><li>如果蠕虫一直为&gt; 1-2分钟的滤纸在该过程期间的任何时刻，添加蠕虫水一滴再水合蜗杆。 注:脱水会增加病毒的易感性翻录受伤。 </li></ol></li><li>重复步骤3.3，直到所有的黑色颜料的组织细胞（色素细胞）的，以及所有的视杯（感光细胞的树突）的白色组织的，被除去。用纸巾擦拭小心的擦拭取出切除组织粘在针的末端。如果双眼睛阿夫拉和灰是期望的，在这一点上烧蚀第二只眼睛。 注:为避免受伤，针可抓住用干净的纸巾针擦拭举行的拇指和食指，然后用另一只手通过擦去拇指和食指拉针进行清洗。可替代地，针可以被擦拭到一个湿纸巾擦拭的表面并检测清洁度。 </li><li> 当完成后，可使用移液管到蜗杆移动到含有新鲜水虫标记的菜。如果分析组数据，放置最多20个蠕虫在一个100毫米的培养皿。如果随着时间的推移在同一个人的跟踪再生，放置1个蜗杆成12孔板的每个孔中。一旦所有蠕虫被转移，通过去除从餐具/孔所有蠕虫水中，并用更多的新鲜水蠕虫更换冲洗蠕虫。 <ol><li>从过滤器除去蠕虫，后送与蜗杆水少量移液管并释放水到所述蠕虫。这应该解除ŧ他蠕虫关闭滤纸，立即被吸进吸管转移。 </li></ol></li><li>孵育约20℃（室温）下避光实验并按照再生过程。 注:蠕虫可被存储在一个温度控制的恒温箱中保持恒定的温度，但是这不是严格必需的。大部分房间的温度只有+ 5°C，这将不会影响该蠕虫的再生能力有所不同。 </li><li>如果需要的话，固定用于免疫组织化学蠕虫（参见图3-4）和/或基因表达的原位杂交分析。不同的固定方法为免疫组织化学涡虫18，19，20和原位杂交21，22，23个协议存在。 </li><li>当实验结束后，sacrifiCE由从培养皿中除去蠕虫水中，并用70％的乙醇代替活蠕虫。蠕虫孵育3-5分钟，检查蠕虫病毒裂解和转灰。 </li><li>发生一次牺牲了正常的废物流中未经处理的蠕虫。避免将活虫体进入环境，特别是非本地物种。 </li></ol>

<ol>
	<li>Gentile, L., Cebria, F., Bartscherer, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+planarian+flatworm:+an+in+vivo+model+for+stem+cell+biology+and+nervous+system+regeneration.">The planarian flatworm: an in vivo model for stem cell biology and nervous system regeneration.</a> Dis Model Mech. 4 (1), 12-19 (2011).</li><li>Elliott, S. A., Sanchez Alvarado, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+history+and+enduring+contributions+of+planarians+to+the+study+of+animal+regeneration.">The history and enduring contributions of planarians to the study of animal regeneration.</a> Wiley Interdiscip Rev Dev Biol. 2 (3), 301-326 (2013).</li><li>Emili Sal&#243;, R. B., Tsonis, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+3.">Chapter 3.</a> Animal Models in Eye Research. , 15-26 (2008).</li><li>Lapan, S. W., Reddien, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=dlx+and+sp6-9+Control+optic+cup+regeneration+in+a+prototypic+eye.">dlx and sp6-9 Control optic cup regeneration in a prototypic eye.</a> PLoS Genet. 7 (8), e1002226 (2011).</li><li>Lapan, S. W., Reddien, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transcriptome+analysis+of+the+planarian+eye+identifies+ovo+as+a+specific+regulator+of+eye+regeneration.">Transcriptome analysis of the planarian eye identifies ovo as a specific regulator of eye regeneration.</a> Cell Rep. 2 (2), 294-307 (2012).</li><li>Inoue, T., 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=Morphological+and+functional+recovery+of+the+planarian+photosensing+system+during+head+regeneration.">Morphological and functional recovery of the planarian photosensing system during head regeneration.</a> Zoolog Sci. 21 (3), 275-283 (2004).</li><li>Pineda, D., 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=The+genetic+network+of+prototypic+planarian+eye+regeneration+is+Pax6+independent.">The genetic network of prototypic planarian eye regeneration is Pax6 independent.</a> Development. 129 (6), 1423-1434 (2002).</li><li>Paskin, T. R., Jellies, J., Bacher, J., Beane, W. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Planarian+Phototactic+Assay+Reveals+Differential+Behavioral+Responses+Based+on+Wavelength.">Planarian Phototactic Assay Reveals Differential Behavioral Responses Based on Wavelength.</a> PLoS One. 9 (12), e114708 (2014).</li><li>Raffa, R. B., Martley, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Amphetamine-induced+increase+in+planarian+locomotor+activity+and+block+by+UV+light.">Amphetamine-induced increase in planarian locomotor activity and block by UV light.</a> Brain Res. 1031 (1), 138-140 (2005).</li><li>Sandmann, T., Vogg, M. C., Owlarn, S., Boutros, M., Bartscherer, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+head-regeneration+transcriptome+of+the+planarian+Schmidtea+mediterranea.">The head-regeneration transcriptome of the planarian Schmidtea mediterranea.</a> Genome Biol. 12 (8), R76 (2011).</li><li>Vasquez-Doorman, C., Petersen, C. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+NuRD+complex+component+p66+suppresses+photoreceptor+neuron+regeneration+in+planarians.">The NuRD complex component p66 suppresses photoreceptor neuron regeneration in planarians.</a> Regeneration (Oxf). 3 (3), 168-178 (2016).</li><li>Deochand, M. E., Birkholz, T. R., Beane, W. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temporal+regulation+of+planarian+eye+regeneration.">Temporal regulation of planarian eye regeneration.</a> Regeneration. 3 (4), 209-221 (2016).</li><li>Sakai, F., Agata, K., Orii, H., Watanabe, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Organization+and+regeneration+ability+of+spontaneous+supernumerary+eyes+in+planarians+-eye+regeneration+field+and+pathway+selection+by+optic+nerves.">Organization and regeneration ability of spontaneous supernumerary eyes in planarians -eye regeneration field and pathway selection by optic nerves.</a> Zoolog Sci. 17 (3), 375-381 (2000).</li><li>Asano, Y., Nakamura, S., Ishida, S., Azuma, K., Shinozawa, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rhodopsin-like+proteins+in+planarian+eye+and+auricle:+detection+and+functional+analysis.">Rhodopsin-like proteins in planarian eye and auricle: detection and functional analysis.</a> J Exp Biol. 201 (Pt 9), 1263-1271 (1998).</li><li>Cross, S. D., 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=Control+of+Maintenance+and+Regeneration+of+Planarian+Eyes+by+ovo.">Control of Maintenance and Regeneration of Planarian Eyes by ovo.</a> Invest Ophthalmol Vis Sci. 56 (12), 7604-7610 (2015).</li><li>Robb, S. M., Gotting, K., Ross, E., Sanchez Alvarado, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SmedGD+2.0:+The+Schmidtea+mediterranea+genome+database.">SmedGD 2.0: The Schmidtea mediterranea genome database.</a> Genesis. 53 (8), 535-546 (2015).</li><li>Robb, S. M., Ross, E., Sanchez Alvarado, ., A, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SmedGD:+the+Schmidtea+mediterranea+genome+database.">SmedGD: the Schmidtea mediterranea genome database.</a> Nucleic Acids Res. 36, D599-D606 (2008).</li><li>Beane, W. S., Tseng, A. S., Morokuma, J., Lemire, J. M., Levin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+planar+cell+polarity+extends+neural+growth+during+regeneration,+homeostasis,+and+development.">Inhibition of planar cell polarity extends neural growth during regeneration, homeostasis, and development.</a> Stem Cells Dev. 21 (12), 2085-2094 (2012).</li><li>Forsthoefel, D. J., Waters, F. A., Newmark, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generation+of+cell+type-specific+monoclonal+antibodies+for+the+planarian+and+optimization+of+sample+processing+for+immunolabeling.">Generation of cell type-specific monoclonal antibodies for the planarian and optimization of sample processing for immunolabeling.</a> BMC Dev Biol. 14, 45 (2014).</li><li>Ross, K. G., 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=Novel+monoclonal+antibodies+to+study+tissue+regeneration+in+planarians.">Novel monoclonal antibodies to study tissue regeneration in planarians.</a> BMC Dev Biol. 15, 2 (2015).</li><li>Cardona, A., Fernandez, J., Solana, J., Romero, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+in+situ+hybridization+protocol+for+planarian+embryos:+monitoring+myosin+heavy+chain+gene+expression.">An in situ hybridization protocol for planarian embryos: monitoring myosin heavy chain gene expression.</a> Dev Genes Evol. 215 (9), 482-488 (2005).</li><li>King, R. S., Newmark, P. 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+situ+hybridization+protocol+for+enhanced+detection+of+gene+expression+in+the+planarian+Schmidtea+mediterranea.">In situ hybridization protocol for enhanced detection of gene expression in the planarian Schmidtea mediterranea.</a> BMC Dev Biol. 13, 8 (2013).</li><li>Pearson, B. 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=Formaldehyde-based+whole-mount+in+situ+hybridization+method+for+planarians.">Formaldehyde-based whole-mount in situ hybridization method for planarians.</a> Dev Dyn. 238 (2), 443-450 (2009).</li></ol>

作者什么都没有透露。

此眼消融技术提高了对当前方法（如打孔）通过排除脑组织和主要限制切除到眼组织中。通过实践，这项技术可以被大多数人来执行，技术人员在显微外科经验不足，但认真的本科生经历。因此建议该技术使用在实验的消融，包括通过免疫组织化学或在眼睛标志物4，5，13 原位杂交（如果可能）完全去除所有眼部?...

涡虫是研究成体干细胞介导再生一个功能强大的模式生物。这些非寄生的扁虫淡水拥有再生的任何及所有丢失的组织，包括他们的中枢神经系统和大脑1的能力。研究早在1700年2，技术在涡领域在过去10 - 15年的进展（如基因组测序， 原位杂交，免疫组织化学，RNA干扰（RNAi），和转录）已经更新了这个历史的模式生物。具体来说，涡虫最近获得了普及作为眼科研究3一个新兴的模式。 涡具有只有两种组织类型，在感光体的神经元和色素细胞原型眼睛;这使得它的眼干细胞群的鉴定和证明，许多相同的基因调节脊椎动物的眼睛去velopment是保守的涡虫4,5。光学杯位于背侧和包括感光体神经元的白色，无颜料的树突和所述半月形黑色素细胞，眼睛经由视交叉支配脑。除了作为用于阐明再生过程6所述的模型，涡虫眼是非常适合于学习的视觉机构7的演变，行为反应的光（涡显示负趋光）8，和行为9的神经学基础。 在涡眼再生大部分已经研究主要有两种情况:作为头再生的一部分以下断头4，图 10和以下只是眼部组织11的切除，12 </SUP&gt;。眼睛上再生涡虫的大多数研究都使用了断头方法，因为它是简单明了。最常见的涡眼切除方法迄今通过打孔器是用细玻璃毛细管13，14，但也有一些研究也已经进行截肢仅次于眼（局部断头）15。然而，所有的这些方法涉及的不仅仅是眼睛许多其他组织（如脑，肠，肾管）的损失，潜在的复杂结果的解释。这里介绍的眼睛消融协议限制切除到更特定于眼内的眼组织（特别排除脑），导致数据。另外，称取7-14天，开始馈送断头蠕虫不同，眼烧蚀蠕虫将消融12的24小时内进料，允许RNAi实验（其中的RNAi经由食物递送）被performeð兼任。 虽然眼睛消融术在技术上是很难比斩首成功执行，涉及眼切除目前的研究尚未包括在他们的程序的详细说明。这个视频文章的目的是为了使研究人员能够一致地删除涡视杯，而不会干扰底层的脑组织和撤除其他一些组织成为可能。此方法可用于单和双眼消融，并适用于广泛的调查。像大多数再生试验中，眼消融非常适合结合两者的药理学和遗传学（RNAi）的屏幕，以及行为研究。这里，我们介绍的方法蠕虫的处理，保持了涡虫的殖民地，和眼睛消融技术本身。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Instant Ocean sea salts</td>
			<td>Spectrum Brands&#160;</td>
			<td>SS15-10</td>
			<td>&#34;10 Gallon&#34; box&#160; (net weight 3 lbs)</td>
		</tr>
		<tr>
			<td>Kimwipes EX-L lint-free tissue wipe</td>
			<td>Kimberly-Clark</td>
			<td>34155</td>
			<td>4.5 x 8.5 in&#160;</td>
		</tr>
		<tr>
			<td>Whatman #2 filter paper</td>
			<td>Sigma&#160;</td>
			<td>WHA1002125</td>
			<td>Circles, 125 mm diameter, white</td>
		</tr>
		<tr>
			<td>Easy Touch Insulin syringe (with needle)</td>
			<td>Pet Health Market</td>
			<td>17175-04</td>
			<td>&#160;U-100 1 cc syringe, 31-gauge 5/16 in needle</td>
		</tr>
		<tr>
			<td>100 mm Petri dish</td>
			<td>VWR</td>
			<td>25384-342</td>
			<td>100 x 15 mm</td>
		</tr>
		<tr>
			<td>60 mm Petri dish</td>
			<td>VWR</td>
			<td>25384-092</td>
			<td>60 x 15 mm</td>
		</tr>
		<tr>
			<td>Dumont #5&#160; forceps&#160;</td>
			<td>Fine Science Tools</td>
			<td>11254-20</td>
			<td>Inox, straight tip , 11 cm</td>
		</tr>
		<tr>
			<td>Transfer pipettes&#160;</td>
			<td>Samco Scientific&#160;</td>
			<td>225</td>
			<td>Graduated, large bulb, 7.5 mL, non sterile</td>
		</tr>
		<tr>
			<td>Parafilm M paraffin film</td>
			<td>Brand&#160;</td>
			<td>701606</td>
			<td>&#160;4 in x 125 ft roll</td>
		</tr>
		<tr>
			<td>12-well untreated tissue culture plate</td>
			<td>VWR</td>
			<td>15705-059</td>
			<td>Untreated, flat bottom, sterile, Falcon brand</td>
		</tr>
		<tr>
			<td>Plastic food containers (for colony)&#160;</td>
			<td>Ziploc</td>
			<td>Large rectangle</td>
			<td>2.25 qt (2.12 L), 10&#34; x 6 -3/4 &#34; x 3 -3/16&#34;&#160;</td>
		</tr>
		<tr>
			<td>Planaria (Girardia tigrina)</td>
			<td>Carolina Biological</td>
			<td>132954</td>
			<td>Sold as &#34;Brown&#34; Planaria; most often they are G. tigrina (aka Dugesia tigrina), but sometimes are G. dorotocephala (aka Dugesia dorotocephala); either will work.</td>
		</tr>
		<tr>
			<td>Planaria (Schmidtea mediterranea)</td>
			<td>n/a</td>
			<td>n/a</td>
			<td>S. mediterranea are not commercially available. At this time animals are only obtainable from laboratories that use them and have extra animals.</td>
		</tr>
		<tr>
			<td>Brown paper towels&#160;</td>
			<td>Grainger</td>
			<td>2U229</td>
			<td>9-3/16 x 9-3/8&#34; 1-Ply Multifold Paper Towel, UNBLEACHED</td>
		</tr>
		<tr>
			<td>Wash bottle (for worm water), optional</td>
			<td>VWR</td>
			<td>16650-275</td>
			<td>Wash Bottles, Low-Density Polyethylene, Wide Mouth, 500 mL</td>
		</tr>
		<tr>
			<td>Anti-synapsin antibody, optional</td>
			<td>Developmental Studies Hybridoma Bank</td>
			<td>3C11</td>
			<td>Supernatant</td>
		</tr>
		<tr>
			<td>Anti-arrestin antibody, optional</td>
			<td>n/a</td>
			<td>n/a</td>
			<td>Not commercially available. Kind gift from Michael Levin, Tufts University</td>
		</tr>
		<tr>
			<td>Nalgene Lowboy carboy with spigot (for storing worm water), optional</td>
			<td>Nalge Nunc International Corporation</td>
			<td>2324-0015</td>
			<td>15 L,&#160; polypropylene, low profile makes it easier to fill plastic colony containers&#160;</td>
		</tr>
		<tr>
			<td>Custom Peltier plate, optional</td>
			<td>Williams Machine, Foxboro, MA&#160;</td>
			<td>n/a</td>
			<td>Design specifics courtesy of Junji Morokuma, Tufts University:&#160; Peltier plate is constructed of a standard thermoelectric heat pump (for example, All Electronics Corp Catalog # PJT-1, 30 mm2).&#160; The square heat pump is covered with a thin mirrored surface, then placed inside a 30 mm2 square hole in a circular plexiglass form (~50 mm in diameter). This form is of similar thickness to the heat pump, and fits flush into a well tooled in the center of a round heat sink (~115 mm in diameter). The form/heat pump is&#160; &#34;anchored&#34; to the sink with silicone base heat sink compound. The leads are threaded through holes drilled through both the form and the the heat sink. The bottom half of the heat sink is tooled into a &#34;foot&#34; that fits into the opening of your microscope&#39;s base plate.&#160;</td>
		</tr>
		<tr>
			<td>DC power source (for Peltier plate), optional</td>
			<td>B &#38; K Precision</td>
			<td>1665</td>
			<td>Regulated Low Voltage DC Power Supply, 1-18 volts (DC), 1-10 amps.</td>
		</tr>
		<tr>
			<td>Other common supplies</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Gloves</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Razor blade&#160;</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting scope with gooseneck lighting</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Chopstick rests, optional</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

surgical ablation assay for studying eye regeneration in planarians

在成体干细胞和再生机制的研究中，涡虫扁虫是在体内模型系统中的缝钉。这是因为在很大程度上他们丰富的多能干细胞群和伤害，这将是灾难性的大多数动物后再生的所有细胞和组织类型的能力。近日，涡已经得到普及，作为眼睛再生的典范。其再生整个眼睛能力（包括两种组织类型:色素细胞和光感受器）允许用于调节视觉系统再生机制的清扫。眼消融具有用于检查眼睛特定的途径和机制，其中最重要的是，再生在很大程度上仅仅局限于眼组织上的其他技术（如断头或打孔）几个优点。这个视频本文的目的是演示如何可靠地除去涡视杯，而不会干扰脑或周围组织。蠕虫和已建立的细胞集落的维护的处理也被描述。这种技术使用一个31克，5 / 16-英寸胰岛素针手术舀出固定化在冷板涡虫的视杯。此方法既包括单，双眼烧蚀，与眼睛在1-2周内再生，从而允许一个宽范围的应用。特别地，该消融技术可以很容易地与为了更好地理解的再生机制及其演进，眼干细胞和它们的维护，和phototaxic行为反应和它们的神经学基础的药理学和遗传学（RNA干扰）屏结合。

对于第1-2小时手术后，动物可能比完整的蠕虫（但是他们仍然会移动）表现出下降的运动。如果需要的话，蠕虫将在外科手术后24个小时内吃（例如，RNAi的馈送）。在之后一段时间内同一个人的眼睛再生时，确保每次取虫的照片既手术（完好）之前，在1个小时后消融（HPA）。 4天后消融（DPA）再生色素细胞应该是可见的，并且通过14 DPA整个眼睛将有完全再生（ 图3A-B...

Watch this Scientific Journal Video about Surgical Ablation Assay for Studying Eye Regeneration in Planarians at JoVE.com

Surgical Ablation Assay for Studying Eye Regeneration in Planarians

该协议示出如何一致切除涡眼（光杯），而不干扰周围的组织。使用胰岛素针和注射器，任一个或两个的眼睛可以被烧蚀以促进调查调节眼再生，再生视觉的演变，和光诱导行为的神经基础的机制。

手术消融分析在研究涡虫再生眼

In the study of adult stem cells and regenerative mechanisms, planarian flatworms are a staple in vivo model system. This is due in large part to their ...

surgical-ablation-assay-for-studying-eye-regeneration-in-planarians

Research

JoVE Journal

Developmental Biology

10.9K 次观看.  Western Michigan University. 该协议示出如何一致切除涡眼（光杯），而不干扰周围的组织。使用胰岛素针和注射器，任一个或两个的眼睛可以被烧蚀以促进调查调节眼再生，再生视觉的演变，和光诱导行为的神经基础的机制。 

视频: Surgical Ablation Assay for Studying Eye Regeneration in Planarians

Hepatocyte-specific Ablation in Zebrafish to Study Biliary-driven Liver Regeneration

The liver has a great capacity to regenerate. Hepatocytes, the parenchymal cells of the liver, can regenerate in one of two ways: hepatocyte- or biliary-driven liver regeneration. In hepatocyte-driven liver regeneration, regenerating hepatocytes are derived from preexisting hepatocytes, whereas, in biliary-driven regeneration, regenerating hepatocytes are derived from biliary epithelial cells (BECs). For hepatocyte-driven liver regeneration, there are excellent rodent models that have significantly contributed to the current understanding of liver regeneration. However, no such rodent model exists for biliary-driven liver regeneration. We recently reported on a zebrafish liver injury model in which BECs extensively give rise to hepatocytes upon severe hepatocyte loss. In this model, hepatocytes are specifically ablated by a pharmacogenetic means. Here we present in detail the methods to ablate hepatocytes and to analyze the BEC-driven liver regeneration process. This hepatocyte-specific ablation model can be further used to discover the underlying molecular and cellular mechanisms of biliary-driven liver regeneration. Moreover, these methods can be applied to chemical screens to identify small molecules that augment or suppress liver regeneration.

To help assess the molecular mechanisms underlying zebrafish biliary-driven liver regeneration, we established a liver injury model in which the nitroreductase-expressing hepatocytes are genetically ablated upon metronidazole treatment. In this protocol, we describe how to adeptly manipulate, monitor and analyze hepatocyte ablation and biliary-driven liver regeneration.

肝细胞特异性消融在斑马鱼研究胆驱动肝再生

The liver has a great capacity to regenerate. Hepatocytes, the parenchymal cells of the liver, can regenerate in one of two ways: hepatocyte- or ...

科研

发育生物学

Isolation of Mouse Endometrial Epithelial and Stromal Cells for In Vitro Decidualization

Decidualization is a progesterone-dependent differentiation process of endometrial stromal cells and is a prerequisite for successful embryo implantation. Although many efforts have been made to reveal the underlying mechanisms of decidualization, the exact signaling between the epithelial cells that are in contact with the embryo and the underlying stromal cells remains poorly understood. Therefore, studying decidualization in a way that takes both the epithelial and stromal cells into account could improve our knowledge about the molecular details of decidualization. For this purpose, in vivo models of artificial decidualization are physiologically the most relevant; however, manipulation of intercellular communication is limited. Currently, in vitro cultures of endometrial stromal cells are being used to investigate the modulation of decidualization by several signaling molecules. Conventionally, human or mouse endometrial stromal cells are used. However, the availability of human samples is very often limited. Furthermore, the use of murine tissues is accompanied with variety in the method of culturing. This study presents a validated and standardized method to obtain pure Endometrial Epithelial Cell (EEC) and Stromal Cell (ESC) cultures using adult intact mice treated with estrogen for three consecutive days. The protocol is optimized to improve the yield, viability, and purity of the cells and was further extended in order to study decidualization in a coculture of EEC and ESC. This model may be suitable to exploit the importance of both cell types in decidualization and to evaluate the contribution of significant signaling molecules secreted by EEC or ESC during the intercellular communication.

Isolation of Mouse Endometrial Epithelial and Stromal Cells for In Vitro Decidualization

This study presents a standardized and validated method for the isolation and culture of primary mouse endometrial stromal and epithelial cells, which can be used in a coculture system to study in vitro decidualization.

鼠标子宫内膜上皮细胞和基质细胞的隔离<em&gt;体外</em&gt;蜕膜

Decidualization is a progesterone-dependent differentiation process of endometrial stromal cells and is a prerequisite for successful embryo implantation. ...

Precise Cellular Ablation Approach for Modeling Acute Kidney Injury in Developing Zebrafish

Acute Kidney Injury (AKI) is a common medical condition with a high mortality rate. With the repair abilities of the kidney, it is possible to restore adequate kidney function after supportive treatment. However, a better understanding of how nephron cell death and repair occur on the cellular level is required to minimize cell death and to enhance the regenerative process. The zebrafish pronephros is a good model system to accomplish this goal because it contains anatomical segments that are similar to the mammalian nephron. Previously, the most common model used to study kidney injury in fish was the pharmacological gentamicin model. However, this model does not allow for precise spatiotemporal control of injury, and hence it is difficult to study cellular and molecular processes involved in kidney repair. To overcome this limitation, this work presents a method through which, in contrast to the gentamicin approach, a specific Green Fuorescent Protein (GFP)-expressing nephron segment can be photoablated using a violet laser light (405 nm). This novel model of AKI provides many advantages that other methods of epithelial injury lack. Its main advantages are the ability to &#34;dial&#34; the level of injury and the precise spatiotemporal control in the robust in vivo animal model. This new method has the potential to significantly advance the level of understanding of kidney injury and repair mechanisms.

This work presents a new in vivo model of segmental kidney injury using kidney GFP transgenic zebrafish. The model allows for the induction of the targeted ablation of kidney epithelial cells to show the cellular mechanisms of nephron injury and repair.

精确的细胞消融方法建模发展中的斑马鱼急性肾损伤

Acute Kidney Injury (AKI) is a common medical condition with a high mortality rate. With the repair abilities of the kidney, it is possible to restore ...

Partial Lobular Hepatectomy: A Surgical Model for Morphologic Liver Regeneration

Morphological organ regeneration following acute tissue loss is common among lower vertebrates, but is rarely observed in mammalian postnatal life. Adult liver regeneration after 70% partial hepatectomy results in hepatocyte hypertrophy with some replication in remaining lobes with restoration of metabolic activity, but with permanent loss of the injured lobe&#39;s morphology and architecture. Here, we detail a new surgical method in the neonate that leaves a physiologic environment conducive to regeneration. This model involves amputation of the left lobe apex and a subsequent conservative management regimen, and lacks the necessity for ligation of major liver vessels or chemical injury, leaving a physiologic environment where regeneration may occur. We extend this protocol to amputations on juvenile (P7-14) mice, during which the injured liver transitions from organ regeneration to compensatory growth by hypertrophy. The presented, brief 30 min protocol provides a framework to study the mechanisms of regeneration, its age-associated decline in mammals, and the characterization of putative hepatic stem or progenitors.

Here, we present a new method for partial resection of the left hepatic lobe in neonatal (day 0) mice. This new protocol is suitable for studying acute liver injury and injury response in the neonatal setting.

部分小叶切除术: 形态学肝再生的外科模型

Morphological organ regeneration following acute tissue loss is common among lower vertebrates, but is rarely observed in mammalian postnatal life. Adult ...

Methods for the Study of Regeneration in Stentor

Cells need to be able to regenerate their parts to recover from external perturbations. The unicellular ciliate Stentor coeruleus is an excellent model organism to study wound healing and subsequent cell regeneration. The Stentor genome became available recently, along with modern molecular biology methods, such as RNAi. These tools make it possible to study single-cell regeneration at the molecular level. The first section of the protocol covers establishing Stentor cell cultures from single cells or cell fragments, along with general guidelines for maintaining Stentor cultures. Culturing Stentor in large quantities allows for the use of valuable tools like biochemistry, sequencing, and mass spectrometry. Subsequent sections of the protocol cover different approaches to inducing regeneration in Stentor. Manually cutting cells with a glass needle allows studying the regeneration of large cell parts, while treating cells with either sucrose or urea allows studying the regeneration of specific structures located at the anterior end of the cell. A method for imaging individual regenerating cells is provided, along with a rubric for staging and analyzing the dynamics of regeneration. The entire process of regeneration is divided in three stages. By visualizing the dynamics of the progression of a population of cells through the stages, the heterogeneity in regeneration timing is demonstrated.

Methods for the Study of Regeneration in Stentor

The giant ciliate, Stentor coeruleus, is an excellent system to study regeneration and wound healing. We present procedures for establishing Stentor cell cultures from single cells or cell fragments, inducing regeneration by cutting cells, chemically inducing the regeneration of membranellar band and oral apparatus, imaging, and analysis of cell regeneration.

Stentor再生的研究方法

Cells need to be able to regenerate their parts to recover from external perturbations. The unicellular ciliate Stentor coeruleus is an excellent model ...

A Simplified and Efficient Method to Isolate Primary Human Keratinocytes from Adult Skin Tissue

Primary human keratinocytes isolated from fresh skin tissues and their expansion in vitro have been widely used for laboratory research and for clinical applications. The conventional isolation method of human keratinocytes involves a two-step sequential enzymatic digestion procedure, which has been proven to be inefficient in generating primary cells from adult tissues due to the low cell recovery rate and reduced cell viability. We recently reported an advanced method to isolate human primary epidermal progenitor cells from skin tissues that utilizes the Rho kinase inhibitor Y-27632 in the medium. Compared with the traditional protocol, this new method is simpler, easier, and less time-consuming, and increases epithelial stem cell yield and enhances their stem cell characteristics. Moreover, the new methodology does not require the separation of the epidermis from the dermis, and, therefore, is suitable for isolating cells from different types of adult tissues. This new isolation method overcomes the major shortcomings of conventional methods and is more suitable for producing large numbers of epidermal cells with high potency both for laboratory and for clinical applications. Here, we describe the new method in detail.

Here we present a protocol to efficiently isolate primary human keratinocytes from adult skin tissues. This method simplifies the conventional procedure by using the ROCK Inhibitor Y-27632 in the inoculation medium to spontaneously separate epidermal cells from dermal cells.

一种简便有效的成人皮肤组织中人角质形成细胞的分离方法

Primary human keratinocytes isolated from fresh skin tissues and their expansion in vitro have been widely used for laboratory research and for clinical ...

An Enhanced Green Fluorescence Protein-based Assay for Studying Neurite Outgrowth in Primary Neurons

Neurite outgrowth is a fundamental event in the formation of the neural circuits during nervous system development. Severe neurite damage and synaptic dysfunction occur in various neurodegenerative diseases and age-related degeneration. Investigation of the mechanisms that regulate neurite outgrowth would not only shed valuable light on brain developmental processes but also on such neurological disorders. Due to the low transfection efficiency, it is currently challenging to study the effect of a specific protein on neurite outgrowth in primary mammalian neurons. Here, we describe a simple method for the investigation of neurite outgrowth by the co-transfection of primary rat cortical neurons with EGFP and a protein of interest (POI). This method allows the identification of POI transfected neurons through the EGFP signal, and thus the effect of the POI on neurite outgrowth can be determined precisely. This EGFP-based assay provides a convenient approach for the investigation of pathways regulating neurite outgrowth.

In this report, we describe a simple protocol for studying neurite outgrowth in embryonic rat cortical neurons by co-transfecting with EGFP and the protein of interest.

增强的绿色荧光蛋白测定，用于研究原神经元的神经质生长

Neurite outgrowth is a fundamental event in the formation of the neural circuits during nervous system development. Severe neurite damage and synaptic ...

Confocal Microscope-Based Laser Ablation and Regeneration Assay in Zebrafish Interneuromast Cells

Hair cells are mechanosensory cells that mediate the sense of hearing. These cells do not regenerate after damage in humans, but they are naturally replenished in non-mammalian vertebrates such as zebrafish. The zebrafish lateral line system is a useful model for characterizing sensory hair cell regeneration. The lateral line is comprised of hair cell-containing organs called neuromasts, which are linked together by a string of interneuromast cells (INMCs). INMCs act as progenitor cells that give rise to new neuromasts during development. INMCs can repair gaps in the lateral line system created by&#160;cell death. A method is described here for selective INMC ablation using a conventional laser-scanning confocal microscope and transgenic fish that express green fluorescent protein in INMCs. Time-lapse microscopy is then used to monitor INMC regeneration and determine the rate of gap closure. This represents an accessible protocol for cell ablation that does not require specialized equipment, such as a high-powered pulsed ultraviolet laser. The ablation protocol may serve broader interests, as it could be useful for the ablation of additional cell types, employing a tool set that is already available to many users. This technique will further enable the characterization of INMC regeneration under different conditions and from different genetic backgrounds, which will advance the understanding of sensory progenitor cell regeneration.

Laser ablation is a widely applicable technology for studying regeneration in biological systems. The presented protocol describes use of a standard laser-scanning confocal microscope for laser ablation and subsequent time-lapse imaging of regenerating interneuromast cells in the zebrafish lateral line.

斑马鱼内胚细胞中基于共体显微镜的激光消融和再生测定

Hair cells are mechanosensory cells that mediate the sense of hearing. These cells do not regenerate after damage in humans, but they are naturally ...

Development of Organoids from Mouse Pituitary as In Vitro Model to Explore Pituitary Stem Cell Biology

The pituitary is the master endocrine gland regulating key physiological processes, including body growth, metabolism, sexual maturation, reproduction, and stress response. More than a decade ago, stem cells were identified in the pituitary gland. However, despite the application of transgenic in vivo approaches, their phenotype, biology, and role remain unclear. To tackle this enigma, a new and innovative organoid in vitro model is developed to deeply unravel pituitary stem cell biology. Organoids represent 3D cell structures that, under defined culture conditions, self-develop from a tissue's (epithelial) stem cells and recapitulate multiple hallmarks of those stem cells and their tissue. It is shown here that mouse pituitary-derived organoids develop from the gland's stem cells and faithfully recapitulate their in vivo phenotypic and functional characteristics. Among others, they reproduce the activation state of the stem cells as in vivo occurring in response to transgenically inflicted local damage. The organoids are long-term expandable while robustly retaining their stemness phenotype. The new research model is highly valuable to decipher the stem cells' phenotype and behavior during key conditions of pituitary remodeling, ranging from neonatal maturation to aging-associated fading, and from healthy to diseased glands. Here, a detailed protocol is presented to establish mouse pituitary-derived organoids, which provide a powerful tool to dive into the yet enigmatic world of pituitary stem cells.

Development of Organoids from Mouse Pituitary as In Vitro Model to Explore Pituitary Stem Cell Biology

The pituitary gland is the key regulator of the body's endocrine system. This article describes the development of organoids from the mouse pituitary as a novel 3D in vitro model to study the gland's stem cell population of which the biology and function remain poorly understood.

从小鼠垂体中开发类器官作为 体外 模型以探索垂体干细胞生物学

The pituitary is the master endocrine gland regulating key physiological processes, including body growth, metabolism, sexual maturation, reproduction, ...

Eye Removal in Living Zebrafish Larvae to Examine Innervation-dependent Growth and Development of the Visual System

Zebrafish exhibit remarkable life-long growth and regenerative abilities. For example, specialized stem cell niches established during embryogenesis support continuous growth of the entire visual system, both in the eye and the brain. Coordinated growth between the retinae and the optic tectum ensures accurate retinotopic mapping as new neurons are added in the eyes and brain. To address whether retinal axons provide crucial information for regulating tectal stem and progenitor cell behaviors such as survival, proliferation, and/or differentiation, it is necessary to be able to compare innervated and denervated tectal lobes within the same animal and across animals. 
Surgical removal of one eye from living larval zebrafish followed by observation of the optic tectum achieves this goal. The accompanying video demonstrates how to anesthetize larvae, electrolytically sharpen tungsten needles, and use them to remove one eye. It next shows how to dissect brains from fixed zebrafish larvae. Finally, the video provides an overview of the protocol for immunohistochemistry and a demonstration of how to mount stained embryos in low-melting-point agarose for microscopy.

The article explains how to surgically remove eyes from living zebrafish larvae as the first step toward investigating how retinal input influences optic tectum growth and development. In addition, the article provides information about larval anesthetization, fixation, and brain dissection, followed by immunohistochemistry and confocal imaging.

活斑马鱼幼虫的眼睛去除以检查视觉系统的神经支配依赖性生长和发育

Zebrafish exhibit remarkable life-long growth and regenerative abilities. For example, specialized stem cell niches established during embryogenesis ...

手术消融分析在研究涡虫再生眼

摘要

探索更多视频

此视频中的章节

肝细胞特异性消融在斑马鱼研究胆驱动肝再生

鼠标子宫内膜上皮细胞和基质细胞的隔离

精确的细胞消融方法建模发展中的斑马鱼急性肾损伤

部分小叶切除术: 形态学肝再生的外科模型

Stentor再生的研究方法

一种简便有效的成人皮肤组织中人角质形成细胞的分离方法

增强的绿色荧光蛋白测定，用于研究原神经元的神经质生长

斑马鱼内胚细胞中基于共体显微镜的激光消融和再生测定

从小鼠垂体中开发类器官作为体外模型以探索垂体干细胞生物学

活斑马鱼幼虫的眼睛去除以检查视觉系统的神经支配依赖性生长和发育

手术消融分析在研究涡虫再生眼

摘要

探索更多视频

此视频中的章节

肝细胞特异性消融在斑马鱼研究胆驱动肝再生

鼠标子宫内膜上皮细胞和基质细胞的隔离

精确的细胞消融方法建模发展中的斑马鱼急性肾损伤

部分小叶切除术: 形态学肝再生的外科模型

Stentor再生的研究方法

一种简便有效的成人皮肤组织中人角质形成细胞的分离方法

增强的绿色荧光蛋白测定，用于研究原神经元的神经质生长

斑马鱼内胚细胞中基于共体显微镜的激光消融和再生测定

从小鼠垂体中开发类器官作为 体外 模型以探索垂体干细胞生物学

活斑马鱼幼虫的眼睛去除以检查视觉系统的神经支配依赖性生长和发育

从小鼠垂体中开发类器官作为体外模型以探索垂体干细胞生物学