Name: mosquito-associated virus isolation from field-collected mosquitoes
Uploaded: 2022-08-31T13:20:00
Description: Watch this Scientific Journal Video about Mosquito-Associated Virus Isolation from Field-Collected Mosquitoes at JoVE.com

这项工作得到了武汉市科技计划项目（2018201261638501）的支持。

1. 蚊虫采样和分类
<ol><li>通过现场的轻型捕蚊器MXA-02或二氧化碳捕蚊器诱捕成年蚊子。</li>
	<li>通过浸入液氮10，11杀死收集的蚊子。通过冷链物流系统将它们运送到实验室12. 注意：干冰主要用于冷链物流系统。</li>
	<li>（可选）如果样品地点靠近实验室，则直接将网捕器中的活蚊子送到实验室，并通过在≤-20°C冷冻30<s...

<ol>
	<li>Xia, H., Wang, Y., Atoni, E., Zhang, B., Yuan, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mosquito-associated+viruses+in+China.">Mosquito-associated viruses in China.</a> Virologica Sinica. 33 (1), 5-20 (2018).</li><li>Atoni, E., 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=A+dataset+of+distribution+and+diversity+of+mosquito-associated+viruses+and+their+mosquito+vectors+in+China.">A dataset of distribution and diversity of mosquito-associated viruses and their mosquito vectors in China.</a> Scientific Data. 7 (1), 342 (2020).</li><li>Atoni, E., 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+discovery+and+global+distribution+of+novel+mosquito-associated+viruses+in+the+last+decade+(2007-2017).">The discovery and global distribution of novel mosquito-associated viruses in the last decade (2007-2017).</a> Reviews in Medical Virology. 29 (6), 2079 (2019).</li><li>Xia, H., 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=Comparative+metagenomic+profiling+of+viromes+associated+with+four+common+mosquito+species+in+China.">Comparative metagenomic profiling of viromes associated with four common mosquito species in China.</a> Virologica Sinica. 33 (1), 59-66 (2018).</li><li>Ong, O. T. W., Skinner, E. B., Johnson, B. J., Old, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mosquito-borne+viruses+and+non-human+vertebrates+in+Australia:+A+review.">Mosquito-borne viruses and non-human vertebrates in Australia: A review.</a> Viruses. 13 (2), 265 (2021).</li><li>Agboli, E., Leggewie, M., Altinli, M., Schnettler, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mosquito-specific+viruses-transmission+and+interaction.">Mosquito-specific viruses-transmission and interaction.</a> Viruses. 11 (9), 873 (2019).</li><li>Halbach, R., Junglen, S., van Rij, 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=Mosquito-specific+and+mosquito-borne+viruses:+evolution,+infection,+and+host+defense.">Mosquito-specific and mosquito-borne viruses: evolution, infection, and host defense.</a> Current Opinion in Insect Science. 22, 16-27 (2017).</li><li>Bolling, B. G., Olea-Popelka, F. J., Eisen, L., Moore, C. G., Blair, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transmission+dynamics+of+an+insect-specific+flavivirus+in+a+naturally+infected+Culex+pipiens+laboratory+colony+and+effects+of+co-infection+on+vector+competence+for+West+Nile+virus.">Transmission dynamics of an insect-specific flavivirus in a naturally infected Culex pipiens laboratory colony and effects of co-infection on vector competence for West Nile virus.</a> Virology. 427 (2), 90-97 (2012).</li><li>Baidaliuk, A., 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=Cell-fusing+agent+virus+reduces+arbovirus+dissemination+in+Aedes+aegypti+mosquitoes+in+vivo.">Cell-fusing agent virus reduces arbovirus dissemination in Aedes aegypti mosquitoes in vivo.</a> Journal of Virology. 93 (18), 00715-00719 (2019).</li><li>Atoni, E., 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=Metagenomic+virome+analysis+of+Culex+mosquitoes+from+Kenya+and+China.">Metagenomic virome analysis of Culex mosquitoes from Kenya and China.</a> Viruses. 10 (1), 30 (2018).</li><li>Xia, H., 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=First+isolation+and+characterization+of+a+group+C+Banna+virus+(BAV)+from+Anopheles+sinensis+mosquitoes+in+Hubei,+China.">First isolation and characterization of a group C Banna virus (BAV) from Anopheles sinensis mosquitoes in Hubei, China.</a> Viruses. 10 (10), 555 (2018).</li><li>Shi, C., 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=Stability+of+the+virome+in+lab-+and+field-collected+Aedes+albopictus+mosquitoes+across+different+developmental+stages+and+possible+core+viruses+in+the+publicly+available+virome+data+of+Aedes+mosquitoes.">Stability of the virome in lab- and field-collected Aedes albopictus mosquitoes across different developmental stages and possible core viruses in the publicly available virome data of Aedes mosquitoes.</a> mSystems. 5 (5), 00640 (2020).</li><li>Zhou, M., Chu, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Handbook for Classification and Identification of Main Vectors. , (2019).</li><li>Wang, 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=Identifying+the+main+mosquito+species+in+China+based+on+DNA+barcoding.">Identifying the main mosquito species in China based on DNA barcoding.</a> Plos One. 7 (10), (2012).</li><li>Ratnasingham, S., Hebert, P. D. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bold:+The+Barcode+of+Life+Data+System+(www.barcodinglife.org).">Bold: The Barcode of Life Data System (www.barcodinglife.org).</a> Molecular Ecology Notes. 7 (3), 355-364 (2007).</li><li>Huang, Y., 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=In+vitro+and+in+vivo+characterization+of+a+new+strain+of+mosquito+Flavivirus+derived+from+Culicoides.">In vitro and in vivo characterization of a new strain of mosquito Flavivirus derived from Culicoides.</a> Viruses. 14 (6), 1298 (2022).</li><li>Zhao, L., 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=Characterization+of+a+novel+Tanay+virus+isolated+from+Anopheles+sinensis+mosquitoes+in+Yunnan,+China.">Characterization of a novel Tanay virus isolated from Anopheles sinensis mosquitoes in Yunnan, China.</a> Frontiers in Microbiology. 10, 1963 (1963).</li><li>Ren, N., 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=Characterization+of+a+novel+reassortment+Tibet+orbivirus+isolated+from+Culicoides+spp.+in+Yunnan,+PR+China.">Characterization of a novel reassortment Tibet orbivirus isolated from Culicoides spp. in Yunnan, PR China.</a> Journal of General Virology. 102 (9), 001645 (2021).</li></ol>

该方法的目的是提供一种使用各种细胞系分离蚊子相关病毒的实用方法。将抗生素 - 抗真菌剂（青霉素 - 链霉素 - 两性霉素）添加到蚊子匀浆的上清液中以避免被细菌或真菌污染至关重要。在田间获得的蚊子和病毒上清液必须在-80°C下冷藏，以避免反复冻融循环。
协议中的另一个关键步骤是研磨。蚊子样本应彻底粉末化并储存在冰上。地面不足的蚊子组织可诱发细胞死亡，?...

蚊子是一组重要的致病性节肢动物媒介。Culicidae1，2家族中大约有3，500种蚊子。高通量测序技术的发展导致在世界不同地区的蚊子中发现了许多新的病毒样序列3。一般来说，这些蚊子相关病毒可分为两大类：MBV和MSV。
MBV是一组不同的病毒，是许多人类或动物疾病的病原体，例如黄热病病毒（YFV）、登革热病毒（DENV）、日本脑炎病毒（JEV）、西尼罗河病毒（WNV）和裂谷热病毒（RVFV）4。它们严重威胁着公众健康，导致全世界人类和动物的严重发病率和死亡率。MBV通过从受感染的蚊子传播到幼稚的宿主，以及从病毒感染的宿主和进食的蚊子的传播，自然地维持不同宿主之间的生命周期5。因此，这些病毒可以在实验室中感染蚊子细胞系和脊椎动物细胞系1。
MSV包括宜昌病毒（YCN）、黄库蚊病毒（CxFV）和朝阳病毒（CHAOV），是昆虫特异性病毒1，6，7的一个亚组。近年来，新型MSV的发现有所增加，其中一些MSV被发现对MBV的传播有影响。例如，CxFV可能是毛库蚊的持续感染，可以在早期阶段抑制西尼罗河病毒的复制8。已发现另一种昆虫特异性黄病毒，细胞融合剂病毒（CFAV）可抑制DENV和寨卡病毒（ZIKV）在埃及伊蚊中的繁殖9。因此，该协议是分离蚊子相关病毒的有用方法，可以帮助进一步研究与蚊子相关的病原体的分布和控制蚊媒疾病的控制。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.22 &#181;m membrane filter</td>
			<td>Millipore</td>
			<td>SLGP033RB</td>
			<td>Polymer films with specific pore ratings.To remove cell debris and bacteria.</td>
		</tr>
		<tr>
			<td>24-well plates</td>
			<td>CORNING</td>
			<td>3524</td>
			<td>Containers for cell</td>
		</tr>
		<tr>
			<td>75 cm2 flasks</td>
			<td>CORNING</td>
			<td>430641</td>
			<td>Containers for cell</td>
		</tr>
		<tr>
			<td>a sterile 2 mL tube with 3 mm ceramic beads</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Antibiotic-Antimycotic</td>
			<td>Gibco</td>
			<td>15240-062</td>
			<td>Antibiotic in the medium to prevent contamination from bacteria and fungi</td>
		</tr>
		<tr>
			<td>Automated nucleic acid extraction system</td>
			<td>NanoMagBio</td>
			<td>S-48</td>
			<td></td>
		</tr>
		<tr>
			<td>BHK-21 cells</td>
			<td>National Virus Resource Center, Wuhan Institute of Virology</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>C6/36 cells</td>
			<td>National Virus Resource Center, Wuhan Institute of Virology</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifugal machine</td>
			<td>Himac</td>
			<td>CF16RN</td>
			<td>Instrument for centrifugation of mosquito samples</td>
		</tr>
		<tr>
			<td>CO2</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#8217;s minimal essential medium (DMEM)</td>
			<td>Gibco</td>
			<td>C11995500BT</td>
			<td>medium for vertebrate cell lines</td>
		</tr>
		<tr>
			<td>Ebinur Lake virus</td>
			<td></td>
			<td></td>
			<td>Cu20-XJ isolation</td>
		</tr>
		<tr>
			<td>Feta Bovine Serum (FBS)</td>
			<td>Gibco</td>
			<td>10099141C</td>
			<td> 
			 
			Provide nutrition for cells</td>
		</tr>
		<tr>
			<td>high-speed low-temperature tissue homogenizer</td>
			<td>servicebio</td>
			<td>KZ-III-F</td>
			<td>Instrument for grinding</td>
		</tr>
		<tr>
			<td>incubator (28 &#176;C)</td>
			<td>Panasonic</td>
			<td>MCO-18AC</td>
			<td>Instrument for cell culture</td>
		</tr>
		<tr>
			<td>incubator (37 &#176;C)</td>
			<td>Panasonic</td>
			<td>MCO-18AC</td>
			<td>Instrument for cell culture</td>
		</tr>
		<tr>
			<td>PCR tube</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>penicillin-streptomycin</td>
			<td>Gibco</td>
			<td>15410-122</td>
			<td>Antibiotic in the medium to prevent contamination from bacteria</td>
		</tr>
		<tr>
			<td>Penicillin-Streptomycin-Amphotericin B Solution</td>
			<td>Gibco</td>
			<td>15240096</td>
			<td></td>
		</tr>
		<tr>
			<td>Refrigerator (-80 &#176;C)</td>
			<td>sanyo</td>
			<td>MDF-U54V</td>
			<td></td>
		</tr>
		<tr>
			<td>Roswell Park Memorial Institute&#160; medium (RPMI)</td>
			<td>Gibco</td>
			<td>C11875500BT</td>
			<td>medium for mosiquto cell lines</td>
		</tr>
		<tr>
			<td>Screw cap storage tubes (2 mL)</td>
			<td>biofil</td>
			<td>&#160;FCT010005</td>
			<td></td>
		</tr>
		<tr>
			<td>sterile pestles</td>
			<td>Tiangen</td>
			<td>OSE-Y004</td>
			<td>Consumables&#160; for grinding</td>
		</tr>
		<tr>
			<td>TGrinder OSE-Y30 electric tissue grinder</td>
			<td>Tiangen</td>
			<td>OSE-Y30</td>
			<td>Instrument for grinding</td>
		</tr>
		<tr>
			<td>The dissecting microscope</td>
			<td>ZEISS</td>
			<td>stemi508</td>
			<td></td>
		</tr>
		<tr>
			<td>the light traps MXA-02</td>
			<td>Maxttrac</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>The mosquito absorbing machine</td>
			<td>Ningbo Bangning</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>The pipette tips</td>
			<td>Axygen</td>
			<td>TF</td>
			<td></td>
		</tr>
		<tr>
			<td>The QIAamp viral RNA mini kit</td>
			<td>QIAGEN</td>
			<td>52906</td>
			<td></td>
		</tr>
		<tr>
			<td>Tweezers</td>
			<td>Dumont</td>
			<td>0203-5-PO</td>
			<td></td>
		</tr>
	</tbody>
</table>

mosquito-associated virus isolation from field-collected mosquitoes

随着测序技术的广泛应用，在包括蚊子在内的节肢动物中发现了许多新型病毒样序列。这些新的蚊媒病毒的两个主要类别是“蚊媒病毒（MBV）”和“蚊媒病毒（MSV）”。这些新型病毒可能对脊椎动物和蚊子都有致病性，或者它们可能只是与蚊子共生。实体病毒对于确认这些病毒的生物学特征至关重要。因此，这里描述了从现场收集的蚊子中分离和扩增病毒的详细方案。首先，制备蚊子样品作为蚊子匀浆的上清液。离心两次后，将上清液接种到蚊子细胞系C6/36或脊椎动物细胞系BHK-21中进行病毒扩增。7天后，收集上清液作为P1上清液并储存在-80°C。 接下来，P1上清液在C6 / 36或BHK-21细胞中再传代两次，同时每天检查细胞状态。当发现细胞的细胞致病作用（CPE）时，收集这些上清液并用于鉴定病毒。该协议是未来研究蚊子相关病毒（包括MBV和MSV）的基础。

用蚊子匀浆（P0）的上清液接种后，C6 / 36细胞表现出广泛的细胞间空间，并且在120小时（图1A）与未接种的细胞（对照）相比观察到脱落的细胞（图1B）同时（图1B）。将BHK-21细胞与P3上清液孵育后，与对照细胞（图1D）相比，在48小时（图1C）在BHK-21细胞中观察到可见的CPE。进行PCR以确定病毒种类。用于检测?...

Watch this Scientific Journal Video about Mosquito-Associated Virus Isolation from Field-Collected Mosquitoes at JoVE.com

Mosquito-Associated Virus Isolation from Field-Collected Mosquitoes

由于测序技术的广泛使用，在蚊子中发现了许多新的病毒样序列。我们提供了一种使用脊椎动物和蚊子细胞系分离和扩增病毒的有效程序，这可能作为未来研究蚊子相关病毒（包括蚊媒和蚊子特异性病毒）的基础。

从田间收集的蚊子中分离出蚊相关病毒

With the broad application of sequencing technologies, many novel virus-like sequences have been discovered in arthropods, including mosquitoes. The two ...

该方案是分离蚊子相关病毒的有用方法，可以帮助进一步研究与蚊子相关的病原体的分布和控制蚊媒疾病的控制。在生物安全2级实验室中使用细线分离病毒更安全，更有效。与其他病毒分离程序不同，例如用不同的液体循环接种红色，该技术不需要动物实验或伦理评估。孵育过程中样品和细胞抽取的污染可能导致实验失败。因此，有必要在培养基中添加2%青霉素-链霉素-两性霉素以防止污染。首先在放置在冰上的两毫升无菌管中加入三毫米陶瓷珠，1.5毫升RPMI培养基补充2%青霉素 - 链霉素 - 两性霉素B溶液，然后在其中转移50只蚊子。接下来，用低温组织均质器在 70 赫兹和 4 摄氏度下研磨 30 秒，使蚊子均质化 3 个周期。将匀浆在15， 000G下在4摄氏度下离心30分钟，并将上清液转移到新管中。为了完全去除蚊子碎片，在4摄氏度下再次以15， 000G离心上清液10分钟。将200微升上清液P0等分到两毫升的螺帽储存管中，并将其储存在零下80摄氏度。对于病毒扩增，使用白纹伊蚊RNAi缺陷蚊细胞系C6 / 36和脊椎动物细胞系，小仓鼠肾或来自75厘米方形烧瓶的BHK-21，并分别将它们接种在24孔板中。将接种的 24 孔板在 28 或 37 摄氏度下过夜。第二天，在显微镜下观察细胞，以确认每个孔中的80%至90%汇合度。准备500毫升补充有抗生素的细胞培养维持培养基。从24孔板中取出细胞培养基，并向每个孔中加入100微升细胞培养维持培养基。然后用100微升蚊子匀浆或P0的上清液接种细胞。将板在28或37摄氏度下孵育60分钟，每15分钟轻轻摇动板以防止细胞干燥。完成后，除去上清液并用600微升细胞培养维持培养基冲洗每个孔，以完全去除碎片。接下来，向每个孔中加入800微升细胞培养维持培养基，然后将板置于28或37摄氏度的5%二氧化碳加湿培养箱中7天。每天在显微镜下监测每个孔的细胞状态。在第七天，收集称为P1的细胞上清液并将其储存在零下80摄氏度。重复该过程以收集P2和P3病毒上清液。由于细胞致病效应或CPE，来自某些孔的细胞死亡并从表面分离。为了极大地扩增病毒从显示CPE效应的孔中收集300至400微升上清液，并将其转移到具有80%至90%细胞汇合度的培养细胞的新六孔板中。最后，收集500微升上清液并将其从六孔板的孔转移到两毫升螺旋盖储存管中，然后将其储存在零下80摄氏度。与未接种或对照细胞相比，用蚊子匀浆P0接种C6 / 36细胞在感染后120小时内在脱落细胞中表现出广泛的细胞间空间。BHK21细胞与P3病毒上清液孵育在感染后48小时显示出可见的CPE，与对照细胞相比，显示细胞死亡并从表面分离。市售通用引物或黄病毒、甲病毒和布尼亚病毒用于生成病毒上清液的PCR产物。布尼亚病毒的PCR产物Ebinur Lake病毒被设定为阳性对照。属于布尼亚病毒的泳道中的PCR扩增禁令和阳性对照强调了上清液中存在布尼亚病毒的可能性。为了提高病毒分离的成功率，在样品运输、分离和研磨过程中保持低温。在均质蚊子和培养物本身时添加抗生素。并在孵育和去除培养基期间快速处理培养物。除此程序外，还可以将研磨溶液注射到循环大鼠或鸡胚中。然而，确认程序会引起伦理问题并增加实验费用。

Research

JoVE Journal

Biology

Dissection of Midgut and Salivary Glands from Ae. aegypti Mosquitoes

从AE肠和唾液腺的解剖。埃及伊蚊蚊

The mosquito midgut and salivary glands are key entry and exit points for pathogens such as Plasmodium parasites and Dengue viruses. This video protocol ...

科研

生物学

Isolation of Genomic DNA from Mouse Tails

从鼠尾基因组DNA提取

Mitochondrial Isolation from Skeletal Muscle

骨骼肌的线粒体分离

Mitochondria are organelles controlling the life and death of the cell. They participate in key metabolic reactions, synthesize most of the ATP, and ...

Isolation of Immune Cells from Primary Tumors

从原发肿瘤的免疫细胞的分离

Tumors create a unique immunosuppressive microenvironment (tumor microenvironment, TME) whereby leukocytes are recruited into the tumor by various ...

The MultiBac Protein Complex Production Platform at the EMBL

在EMBL的MultiBac蛋白质复杂的生产平台

Proteomics  research revealed the impressive complexity of eukaryotic proteomes in  unprecedented detail. It is now a commonly accepted notion that ...

Isolation of Myofibroblasts from Mouse and Human Esophagus

从小鼠和人类食管隔离肌纤维母细胞

Murine and human esophageal myofibroblasts are generated via enzymatic digestion. Neonate (8-12 day old) murine esophagus is harvested, minced, washed, ...

A Multi-detection Assay for Malaria Transmitting Mosquitoes

一种多检测试验为疟疾蚊子发射

The Anopheles gambiae species complex includes the major malaria transmitting mosquitoes in Africa. Because these species are of such medical importance, ...

Protocols for Testing the Toxicity of Novel Insecticidal Chemistries to Mosquitoes

新型杀虫化学对蚊子毒性测试规程

New classes of insecticides with novel modes of action are needed to control insecticide resistant populations of mosquitoes that transmit diseases such ...

Forced Salivation As a Method to Analyze Vector Competence of Mosquitoes

强迫流涎法分析蚊虫的载体能力

Vector competence is defined as the potential of a mosquito species to transmit a mosquito-borne virus (mobovirus) to a vertebrate host. Viable virus ...

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 ...