Name: retro-orbital injection in adult zebrafish
Uploaded: 2009-12-07T18:43:00
Description: Watch this Scientific Journal Video about Retro-orbital Injection in Adult Zebrafish at JoVE.com

这项工作是霍华德休斯医学研究所和美国国立卫生研究院研究胚胎造血授予的支持。

第1部分。注射材料的制备<ol><li> TG（ 珠 ：GFP）细胞： </li><li>出血成人捐助鱼用10ul枪头涂肝素（1unit/ul）穿刺背后刺鱼。 </li><li>吸红血细胞和免除在细胞液（0.9倍PBS + 5％FBS + 1％笔/链球菌）。 </li><li>超过40uM网过滤细胞悬液，并确定使用血球细胞作为先前由勒布朗等浓度。 </li><li>降速和重新悬浮细胞缓冲所需浓度，这样最后的注射量不超过5ul。在这里，我们注入1.5-2万个细胞/收件人。 </l...

<ol>
	<li>Steel, C. D., Stephens, A. L., Hahto, S. M., Singletary, S. J., Ciavarra, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+the+lateral+tail+vein+and+the+retro-orbital+venous+sinus+as+route+of+intravenous+drug+delivery+in+a+transgenic+mouse+model.">Comparison of the lateral tail vein and the retro-orbital venous sinus as route of intravenous drug delivery in a transgenic mouse model.</a> Lab Animal. 37, 26-32 (2008).</li><li>Pinkerton, W., Webber, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+method+of+injecting+small+laboratory+animals+by+the+ophthalmic+plexus+route.">A method of injecting small laboratory animals by the ophthalmic plexus route.</a> Proc. Soc. Exp. Bio. Med. 116, 959-961 (1964).</li><li>White, R. M., Sessa, A. S., Burke, C., Bowman, T., LeBlanc, J., Ceol, C., Bourque, C., Dovey, M., Goessling, W., Burns, C. E., Zon, L. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transparent+adult+zebrafish+as+a+tool+for+in+vivo+transplantation+analysis.">Transparent adult zebrafish as a tool for in vivo transplantation analysis.</a> Cell Stem Cell. 2, 183-189 (2008).</li><li>LeBlanc, J., Bowman, T. V., Zon, L. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transplantation+of+whole+kidney+marrow+in+adult+zebrafish.">Transplantation of whole kidney marrow in adult zebrafish.</a> J. Vis. Exp. , (2007).</li></ol>

在成年斑马鱼的眼窝注射使注射材料交付的效率最高，血液中的死亡率与发病率最低。由于注射部位的性质，出现穿刺愈合快，减少发生感染和出血量。该网站甚至可能被多次注入在一个短的时间内每天注射药物，例如。 在这种鱼是简单的浸泡和浸泡在含有药物的利息的水成鱼的传统治疗方法，必须购买大量毒品。然而，如果药物直接注入动物的血液，一个更小的数量是需?...

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<td>Heparin sodium salt from porcine intestinal mucosa</td>
<td>&nbsp;</td>
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<td>Preweighed vial of 300 USP units</td>
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<td>Tricaine-S</td>
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<td>Western Chemical</td>
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<td>Dulbecco&#x2019;s Phosphate Buffered Saline</td>
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<td>Dextran, Texas Red&reg;</td>
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0.17 mM Kcl 
0.33 mM CaCl2 
0.33 mM MgSO2
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retro-orbital injection in adult zebrafish

整体动物药物治疗是一种必不可少的工具，在任何药理和化学遗传研究的模型系统。静脉（IV）注射液往往是最有效和无创性的形式利益的代理人交付。在斑马鱼（<em&gt;斑马鱼</em&gt;），四，注射药物一直是因为小血管直径挑战。这也证明了在hematopoeitic干细胞移植过程中的细胞注射的重大障碍。从历史上看，注射到血液中，直接通过心脏。然而，这种心脏内的过程有一个非常高的死亡率，心往往是在离开容易发生感染，大量失血或致命的器官损害的鱼注射刺破。我们用鼠标的经验，我们开发了一个新的注射过程，在斑马鱼注射部位在背后的眼睛，眼窝静脉窦。这种复古的轨道（RO）注射技术已成功地采用了在成鱼注射药物以及整个肾骨髓细胞移植。 RO注入了一个比传统的心脏内注射的死亡率要低得多。鱼类，注入复古轨道往往出血少的注射后，在风险的伤害要低得多的一个主要器官如心。此外，正确执行时，注入的细胞和/或药物迅速进入血液，使化合物，全鱼和肾细胞施加影响，以自己的优势轻松地回家。因此，这种新的注射技术，最大限度地降低了死亡率，同时允许有效地提供材料到成鱼的血液。在这里，我们举例说明这种技术由眼窝TG注射（<em&gt;珠</em&gt;：GFP）的细胞转化为成人<em&gt;卡斯帕尔</em&gt;鱼以及一个红色荧光染料注射液（右旋糖酐，德克萨斯红<em&gt;</em&gt;）到成年<em&gt;卡斯帕尔</em&gt;鱼。然后，我们整个动物荧光显微镜可视化成功的注射。

Watch this Scientific Journal Video about Retro-orbital Injection in Adult Zebrafish at JoVE.com

Retro-orbital Injection in Adult Zebrafish

在这里，我们展示如何在成年斑马鱼眼窝注射。

在成年斑马鱼的眼窝射出

Drug treatment of whole animals is an essential tool in any model system for pharmacological and chemical genetic studies. Intravenous (IV) injection is ...

Research

JoVE Journal

Biology

Transplantation of Whole Kidney Marrow in Adult Zebrafish

在成年斑马鱼的全肾骨髓移植

Hematopoietic stem cells (HSC) are a rare population of pluripotent cells that maintain all the differentiated blood lineages throughout the life of an ...

科研

生物学

Zebrafish Brain Ventricle Injection

斑马鱼脑室注射液

Proper brain ventricle formation during embryonic brain development is required for normal brain function.  Brain ventricles are the highly conserved ...

Dissection of Organs from the Adult Zebrafish

剖析器官的成年斑马鱼

Over the last 20 years, the zebrafish has become a powerful model organism for understanding vertebrate development and disease.  Although experimental ...

Intraperitoneal Injection into Adult Zebrafish

腹腔注射到成年斑马鱼

A convenient method for chemically treating zebrafish is to introduce the reagent into the tank water, where it will be taken up by the fish. However, ...

Dissection of the Adult Zebrafish Kidney

成年斑马鱼肾脏的解剖

Researchers working in the burgeoning field of adult stem cell biology seek to understand the signals that regulate the behavior and function of stem ...

Heart Dissection in Larval, Juvenile and Adult Zebrafish, Danio rerio

Heart Dissection in Larval, Juvenile and Adult Zebrafish, Danio rerio

幼虫，少年和成年斑马鱼的心脏解剖，斑马鱼</em

Zebrafish have become a beneficial and practical model organism for the study of embryonic heart development (see recent reviews1-6), however, work ...

In vivo Electroporation of Morpholinos into the Adult Zebrafish Retina

In vivo Electroporation of Morpholinos into the Adult Zebrafish Retina

在体内电穿孔Morpholinos到成年斑马鱼视网膜

Many devastating inherited eye diseases result in progressive and irreversible blindness because humans cannot regenerate dying or diseased retinal ...

In vivo Electroporation of Morpholinos into the Regenerating Adult Zebrafish Tail Fin

In vivo Electroporation of Morpholinos into the Regenerating Adult Zebrafish Tail Fin

再生的成年斑马鱼尾鳍在吗啉代入体内电穿孔

Certain species of urodeles and teleost fish can regenerate their tissues. Zebrafish have become a widely used model to study the spontaneous regeneration ...

Blood Collection for Biochemical Analysis in Adult Zebrafish

在成年斑马鱼的生化分析采血车

The zebrafish has been used as an animal model for studies of several human diseases. It can serve as a powerful preclinical platform for studies of ...

Gavaging Adult Zebrafish

Gavaging成年斑马鱼

The zebrafish has become an important in vivo model in biomedical research. Effective methods must be developed and utilized to deliver compounds or ...