Name: obtaining hemocytes from the hawaiian bobtail squid euprymna scolopes and observing their adherence to symbiotic and non-symbiotic bacteria
Uploaded: 2010-02-11T04:52:00
Description: Watch this Scientific Journal Video about Obtaining Hemocytes from the Hawaiian Bobtail Squid Euprymna scolopes and Observing their Adherence to Symbiotic and Non-Symbiotic Bacteria at JoVE.com

资金来源：康涅狄格州大学研究基金会和分子和细胞生物学系到SVN，西格玛郗赠款援助的研究和安东尼H.和Majorie R. 普罗迪研究生教育奖学金，以AJC

<ol><li>准备500 0.22微米的过滤器消毒的人工海水MLS（FSW;盐度35个百分点）。 0.22微米微米的过滤器过滤器通过人工或自然海水去除微粒和细菌。 </li><li>麻醉放置在2％的乙醇解决方案，在FSW一个成年人夏威夷短尾鱿鱼（Euprymna scolopes） 。约10分钟，放置在麻醉的动物。鱿鱼将停止游泳，并不会积极响应触摸。持续呼吸，表明地幔的运动，和色素细胞活动仍然应该遵守。 </li><li>面临的一个标准蜡剥离托盘腹朝上?...

<ol>
	<li>Koropatnick, T. A., Kimbell, J. R., McFall-Ngai, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Responses+of+host+hemocytes+during+the+initiation+of+the+squid-Vibrio+symbiosis.">Responses of host hemocytes during the initiation of the squid-Vibrio symbiosis.</a> Biol. Bull. 212, 29-39 (2007).</li><li>McFall-Ngai, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unseen+forces:+the+influence+of+bacteria+on+animal+development.">Unseen forces: the influence of bacteria on animal development.</a> Dev. Biol. 242, 1-14 (2002).</li><li>McFall-Ngai, M. J., Ruby, E. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Symbiont+recognition+and+subsequent+morphogenesis+as+early+events+in+an+animal-bacterial+mutualism.">Symbiont recognition and subsequent morphogenesis as early events in an animal-bacterial mutualism.</a> Science. 254, 1491-1494 (1991).</li><li>Nyholm, S. V., Stewart, J. J., Ruby, E. G., McFall-Ngai, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recognition+between+symbiotic+Vibrio+fischeri+and+the+haemocytes+of+Euprymna+scolopes.">Recognition between symbiotic Vibrio fischeri and the haemocytes of Euprymna scolopes.</a> Environ. Microbiol. 11, 483-493 (2009).</li><li>Nyholm, S. V., McFall-Ngai, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+winnowing:+establishing+the+squid-Vibrio+symbiosis.">The winnowing: establishing the squid-Vibrio symbiosis.</a> Nat. Rev. Microbiol. 2, 632-642 (2004).</li><li>Nyholm, S. V., Stabb, E. V., Ruby, E. G., McFall-Ngai, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishment+of+an+animal-bacterial+association:+recruiting+symbiotic+Vibrios+from+the+environment.">Establishment of an animal-bacterial association: recruiting symbiotic Vibrios from the environment.</a> Proc. Natl. Acad. Sci. USA. 97, 10231-10235 (2000).</li><li>Nyholm, S. V., McFall-Ngai, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sampling+the+light-organ+microenvironment+of+Euprymna+scolopes:+description+of+a+population+of+host+cells+in+association+with+the+bacterial+symbiont+Vibrio+fischeri.">Sampling the light-organ microenvironment of Euprymna scolopes: description of a population of host cells in association with the bacterial symbiont Vibrio fischeri.</a> Biol Bull. 195, 89-97 (1998).</li><li>Visick, K. L., Ruby, E. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vibrio+fischeri+and+its+host:+it+takes+two+to+tango.">Vibrio fischeri and its host: it takes two to tango.</a> Curr. Opin. Microbiol. 9, 632-638 (2006).</li></ol>

有关免疫系统的作用，在调解有益的细菌和主机之间的分子信号研究，近年来，我们与他们的菌群真核生物的共同进化理解作出了重大贡献。鱿鱼/弧菌系统本身已经证明了作为一个听话的模型系统回答在这一领域的根本性问题2,3,5,6,8。鱿鱼Euprymna scolopes光器官允许殖民统治完全由发光弧菌鲵 。由于房子的组织，保持与海水接触的细菌，鱿鱼，必须不仅促进具体的共生还要继?...

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<td>Lab-Tek chambered #1.0 borosilicate coverglass system (8-chambers)</td>
<td>&nbsp;</td>
<td>Thermo-Fisher Scientific</td>
<td>155411</td>
<td>&nbsp;</td>
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<td>26.5 G 1ml latex-free insulin syringe</td>
<td>&nbsp;</td>
<td>Becton, Dickinson and Company</td>
<td>C34551</td>
<td>&nbsp;</td>
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<td>Cell Tracker Orange</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>C34551</td>
<td>&nbsp;</td>
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<td>SteREO Discovery V20 Microscope</td>
<td>&nbsp;</td>
<td>Zeiss</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>SP2 Confocal Microscope</td>
<td>&nbsp;</td>
<td>Leica</td>
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obtaining hemocytes from the hawaiian bobtail squid euprymna scolopes and observing their adherence to symbiotic and non-symbiotic bacteria

有关免疫系统的作用，在调解有益的细菌和主机之间的分子信号研究，近年来，我们与他们的菌群真核生物的共同进化理解作出了重大贡献。夏威夷短尾鱿鱼之间的共生协会，<em&gt; Euprymna scolopes</em&gt;和发光细菌<em&gt;弧菌鲵</em&gt;已被利用作为一个模型系统了解动物发育有益细菌的影响。最近的研究表明，血细胞巨噬细胞样，唯一的鱿鱼主机的先天免疫系统的细胞成分，可能发挥了重要作用，在调解这个协会的建立和维护。该协议将展示如何获得血细胞从<em&gt; E。 scolopes</em然后用这些细胞的细菌结合实验。成人鱿鱼先麻醉前用注射器收集主要头侧血管血淋巴。主机的血细胞，提取的血淋巴中，坚持腔盖玻片，然后接触到绿色荧光蛋白标记的共生<em&gt;弧菌鲵</em&gt;和非共生<em&gt;哈维氏弧菌</em&gt;。血细胞用荧光染料（细胞跟踪橙色，Invitrogen公司），然后counterstained可视化使用荧光显微镜。

Watch this Scientific Journal Video about Obtaining Hemocytes from the Hawaiian Bobtail Squid Euprymna scolopes and Observing their Adherence to Symbiotic and Non-Symbiotic Bacteria at JoVE.com

Obtaining Hemocytes from the Hawaiian Bobtail Squid Euprymna scolopes and Observing their Adherence to Symbiotic and Non-Symbiotic Bacteria

该视频将演示如何从夏威夷短尾乌贼的血细胞（血细胞），<em&gt; Euprymna scolopes</em&gt;细胞生物和细菌粘附实验使用。血细胞将用荧光染料染色和GFP标记的细菌接触到。

获得血细胞从夏威夷短尾鱿鱼 Euprymna scolopes和观察他们坚持共生和非共生细菌

Studies concerning the role of the immune system in mediating molecular signaling between beneficial bacteria and their hosts have, in recent years, made ...

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JoVE Journal

Biology

Obtaining Hemocytes from the Hawaiian Bobtail Squid Euprymna scolopes and Observing their Adherence to Symbiotic and Non-Symbiotic Bacteria

Obtaining Eggs from Xenopus laevis Females

从非洲爪蟾女性获得卵子

The eggs of Xenopus laevis intact, lysed, and/or fractionated are useful for a wide variety of experiments. This protocol shows how to induce egg laying, ...

科研

生物学

Obtaining High Quality RNA from Single Cell Populations in Human Postmortem Brain Tissue

人死后脑组织中的单细胞群获得高品质的RNA

We proposed to investigate the gray matter reduction in the superior temporal gyrus seen in schizophrenia patients, by interrogating gene expression ...

Formulations for Freeze-drying of Bacteria and Their Influence on Cell Survival

冷冻干燥细菌及其影响细胞生存的配方

Cellular water can be removed to reversibly inactivate microorganisms to facilitate storage. One such method of removal is freeze-drying, which is ...

A Simple Protocol for Extracting Hemocytes from Wild Caterpillars

一个简单的协议提取血细胞从野生毛毛虫

Insect hemocytes (equivalent to mammalian white blood cells) play an important role in several physiological processes throughout an insect's life cycle ...

Isolation and Culture of Individual Myofibers and their Satellite Cells from Adult Skeletal Muscle

单个肌纤维和成人骨骼肌卫星细胞的分离和培养

Muscle  regeneration in the adult is performed by resident stem cells called satellite  cells. Satellite cells are defined by  their position between the ...

Collection, Isolation and Enrichment of Naturally Occurring Magnetotactic Bacteria from the Environment

从环境中自然产生的趋磁细菌的采集，分离和富集

Magnetotactic bacteria (MTB) are aquatic microorganisms that were first notably described in 19751 from sediment samples collected in salt marshes of ...

Obtaining Specimens with Slowed, Accelerated and Reversed Aging in the Honey Bee Model

获取标本减慢，加速和反老化的蜜蜂型号

Societies of highly social animals feature vast lifespan differences between closely related individuals. Among social insects, the honey bee is the best ...

A Method for Extracting Pigments from Squid Doryteuthis pealeii

A Method for Extracting Pigments from Squid Doryteuthis pealeii

一种从鱿鱼提取色素的方法<em&gt; Doryteuthis pealeii</em

Cephalopods can undergo rapid and adaptive changes in dermal coloration for sensing, communication, defense, and reproduction purposes. These capabilities ...

Split Green Fluorescent Protein System to Visualize Effectors Delivered from Bacteria During Infection

分离绿色荧光蛋白系统可视化细菌在感染过程中传递的效应

Bacteria, one of the most important causative agents of various plant diseases, secrete a set of effector proteins into the host plant cell to subvert the ...

Isolation of High Quality Murine Atrial and Ventricular Myocytes for Simultaneous Measurements of Ca2+ Transients and L-Type Calcium Current

Isolation of High Quality Murine Atrial and Ventricular Myocytes for Simultaneous Measurements of Ca2+ Transients and L-Type Calcium Current

分离高质量鼠心房和心室肌细胞，同时测量 Ca2+ 瞬变和 L 型钙电流

Mouse models play a crucial role in arrhythmia research and allow studying key mechanisms of arrhythmogenesis including altered ion channel function and ...