This research was possible through funding from the University of Neuch&#226;tel. We would like to thank all the students that helped in improving this technique, namely our colleague Kevin Thievent. We would also like to thank the members of the Thomas Lab for making their laboratory available. We would like to thank Janet Teeple for her help with mosquito rearing. We would also like to thanks Loyal Hall in the laboratory of Pr. Tom Baker for his help in the preparation of the micro capillary glass tubes.

1.注射和解剖盐溶液<ol><li>通过加入氯化钠，氯化钾，及氯化钙2到蒸馏水在pH = 6.8，得到1.3毫摩尔NaCl，0.5mM的KCl和0.2mM的氯化钙制备的盐溶液。 </li><li>将1ml的0.1％甲基绿溶液添加到99毫升计盐溶液的着色透明珠。这是0.001％甲基绿"注射液"。 </li><li>然后，加入5毫升0.1％甲基绿溶液至45毫升生理盐水溶液。这个0.01％甲基绿"夹层溶液"是甲基绿10倍以上的浓缩，以促进珠粒在解剖的着色和它们的观察。 注意:如果需要过滤消毒的缓冲区。 </li></ol> 2.毛细准备<ol><li>热拉微细玻璃管以获得非常细的尖端比最小的小珠（φ= 40微米）42稍大。 </li><li>打开和通过打破用镊子尖或锯尖断djust每个毛细管。 </li></ol> 3.饲养蚊子<ol><li>后方冈比亚按蚊在26±1℃，70±5％相对湿度，和12:12小时的光:暗周期。保持成年女性，让他们获得了10％的糖溶液。 </li></ol> 4.用毛细管珠选择<ol><li>在5cm的培养皿中倒入0.009克的带负电荷的小球（40-120微米直径），并加入5毫升注射液30分钟提前。 注意:我们的目标是让染料着色珠，使珠选择用于注射的更容易。高压灭菌珠子，如果他们不进来的无菌罐子。 </li><li>使用安装在毛细吸管灯泡的毛细管，直观地选择一个双目显微镜下在0.1微升的盐溶液中的最小珠。 注意:当同时接种几个珠，0.1微升的总体积选择三个珠。</li></ol> 5.蚊子处理和接种<ol><li>使用昆虫吸引器，将每个蚊子在50毫升管中。 </li><li>通过将管在碎冰（2-5分钟）简要地冷却各蚊子。 </li><li>放置在其右侧蚊子的双目显微镜下。 </li><li>下一个立体显微镜，通过经由胸腔角质层的左侧牢牢刺穿，然后注入液和珠接种与毛细蚊子。 注意:小心不要握住毛细管作为垂直于在蚊子可能损坏飞行肌肉。 </li><li>取下毛细管。 </li><li>使用轻量级昆虫学镊子，将每个女性在覆盖着蚊帐180毫升的塑料杯中分别;让他们获得了10％的糖溶液。 注:警告是必需的与糖的溶液，作为蚊可能会卡在糖溶液滴。 </li></ol> 6.蚊子解剖和珠摄影<ol><li>检查蚊子是活在接种后24小时。如果是这样，检查他们的条件，只保留那些能飞的蚊子。 </li><li>冻结这些蚊子在-20℃。 注:他们可以解剖前几天保持到几个月，如果需要的话。 </li><li>在32X倍的双目显微镜下取出用镊子蚊子的翅膀。固定机翼上用透明胶带载玻片上。写下用于每个蚊子的代码。 </li><li>把玻璃载片在一台扫描仪，并在1200 dpi的扫描。 注意:检查代码是否可读每一个人蚊子。 </li><li>在玻璃显微镜载片，并在夹层溶液，使用镊子胸廓从头部和腹部分离。 </li><li>用镊子打开胸腔检索珠。 注:珠子不移动到腹部，不与彼此接触，其中大部分在叔自由浮动他的血淋巴。要小心，不要中断与镊子珠，作为非珠白化可能很难找到。等待1〜5分钟，为珠，以获得颜色。 </li><li>转移珠在显微镜载玻片剥离液液滴，拍摄照片前。 注:无需洗珠。 </li><li>使用装备有相机的显微镜，采取各胎圈的数字图像在一个标准的照明设置和放大400倍。 注意:不要改变不同的图片之间的照明，使它们的比较。 </li></ol> 7.图像分析:珠黑化<ol><li>打开的图像分析软件。点击"文件"，然后点击"打开"，找到珠的形象。 </li><li>点击"分析"。点击"设置测量。"勾选在"测量设置"窗口中的"平均灰度值"框。 </li><li>点击圆圈选择工具图标。选择珠周长;留在方珠，避免周围的发光光环。 </li><li>点击"分析"菜单上。点击"措施"。获得胎圈的横截面面积的平均灰度值和大小。 <ol><li>要获得每个珠子的黑化值，使用计算器或计算器软件执行以下操作:"256 - 平均灰度值。"黑化的措施是0和256之间的值，其中0表示完全白色图像。 </li></ol></li></ol>

<ol>
	<li>Dong, Y., Aguilar, R., Xi, Z., Warr, E., Mongin, E., Dimopoulos, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anopheles+gambiae+immune+responses+to+human+and+rodent+Plasmodium+parasite+species.">Anopheles gambiae immune responses to human and rodent Plasmodium parasite species.</a> PLoS pathog. 2 (6), e52 (2006).</li><li>Sadd, B. M., Schmid-Hempel, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PERSPECTIVE:+Principles+of+ecological+immunology.">PERSPECTIVE: Principles of ecological immunology.</a> Evolutionary Appl. 2 (1), 113-121 (2008).</li><li>Crompton, P. D., Moebius, 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=Malaria+Immunity+in+Man+and+Mosquito:+Insights+into+Unsolved+Mysteries+of+a+Deadly+Infectious+Disease*.">Malaria Immunity in Man and Mosquito: Insights into Unsolved Mysteries of a Deadly Infectious Disease*.</a> Annu Rev Immuno. 32 (1), 157-187 (2014).</li><li>Schmid-Hempel, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=EVOLUTIONARY+ECOLOGY+OF+INSECT+IMMUNE+DEFENSES.">EVOLUTIONARY ECOLOGY OF INSECT IMMUNE DEFENSES.</a> Annu Rev Entomol. 50 (1), 529-551 (2005).</li><li>Hillyer, J. F., Schmidt, S. L., Christensen, B. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+phagocytosis+and+melanization+of+bacteria+and+Plasmodium+sporozoites+by+hemocytes+of+the+mosquito+Aedes+aegypti.">Rapid phagocytosis and melanization of bacteria and Plasmodium sporozoites by hemocytes of the mosquito Aedes aegypti.</a> J parasito. 89 (1), 62-69 (2003).</li><li>Carissimo, G., Pondeville, 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=Antiviral+immunity+of+Anopheles+gambiae+is+highly+compartmentalized,+with+distinct+roles+for+RNA+interference+and+gut+microbiota.">Antiviral immunity of Anopheles gambiae is highly compartmentalized, with distinct roles for RNA interference and gut microbiota.</a> PNAS. 112 (2), E176-E185 (2015).</li><li>Christensen, B. M., Li, J., Chen, C. -. C., Nappi, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Melanization+immune+responses+in+mosquito+vectors.">Melanization immune responses in mosquito vectors.</a> Trends parasito. 21 (4), 192-199 (2005).</li><li>Collins, F., Sakai, R., 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=Genetic+selection+of+a+Plasmodium-refractory+strain+of+the+malaria+vector+Anopheles+gambiae.">Genetic selection of a Plasmodium-refractory strain of the malaria vector Anopheles gambiae.</a> Science. 234 (4776), 607-610 (1986).</li><li>Warr, E., Lambrechts, L., Koella, J. C., Bourgouin, C., Dimopoulos, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anopheles+gambiae+immune+responses+to+Sephadex+beads:+Involvement+of+anti-Plasmodium+factors+in+regulating+melanization.">Anopheles gambiae immune responses to Sephadex beads: Involvement of anti-Plasmodium factors in regulating melanization.</a> Insect Biochem Molec. 36 (10), 769-778 (2006).</li><li>Fuchs, S., Behrends, V., Bundy, J. G., Crisanti, A., Nolan, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phenylalanine+metabolism+regulates+reproduction+and+parasite+melanization+in+the+malaria+mosquito.">Phenylalanine metabolism regulates reproduction and parasite melanization in the malaria mosquito.</a> PloS one. 9 (1), e84865 (2014).</li><li>Cerenius, L., S&#246;derh&#228;ll, 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+prophenoloxidase-activating+system+in+invertebrates.">The prophenoloxidase-activating system in invertebrates.</a> Immuno Rev. 198, 116-126 (2004).</li><li>Cerenius, L., Lee, B. L., S&#246;derh&#228;ll, 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+proPO-system:+pros+and+cons+for+its+role+in+invertebrate+immunity.">The proPO-system: pros and cons for its role in invertebrate immunity.</a> Trend Immuno. 29 (6), 263-271 (2008).</li><li>Moret, Y., Schmid-Hempel, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Survival for immunity: the price of immune system activation for bumblebee workers. , 1166-1168 (2000).</li><li>Suwanchaichinda, C., Paskewitz, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+Larval+Nutrition,+Adult+Body+Size,+and+Adult+Temperature+on+the+Ability+of+Anopheles+gambiae(Diptera:+Culicidae)+to+Melanize+Sephadex+Beads.">Effects of Larval Nutrition, Adult Body Size, and Adult Temperature on the Ability of Anopheles gambiae(Diptera: Culicidae) to Melanize Sephadex Beads.</a> J Med Entomol. 35 (2), 157-161 (1998).</li><li>Chun, J., Riehle, M., Paskewitz, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+Mosquito+Age+and+Reproductive+Status+on+Melanization+of+Sephadex+Beads+in+Plasmodium-Refractory+and+-Susceptible+Strains+of+Anopheles+gambiae.">Effect of Mosquito Age and Reproductive Status on Melanization of Sephadex Beads in Plasmodium-Refractory and -Susceptible Strains of Anopheles gambiae.</a> J Invertebr Pathol. 66 (1), 11-17 (1995).</li><li>Schwartz, A., Koella, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Melanization+of+Plasmodium+falciparum+and+C-25+Sephadex+Beads+by+Field-Caught+Anopheles+gambiae+(Diptera:+Culicidae)+from+Southern+Tanzania.">Melanization of Plasmodium falciparum and C-25 Sephadex Beads by Field-Caught Anopheles gambiae (Diptera: Culicidae) from Southern Tanzania.</a> J Med Entomol. 39 (1), 84-88 (2002).</li><li>Zahedi, M., Denham, D. A., Ham, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Encapsulation+and+melanization+responses+of+Armigeres+subalbatus+against+inoculated+Sephadex+beads.">Encapsulation and melanization responses of Armigeres subalbatus against inoculated Sephadex beads.</a> J Invertebr Pathol. 59 (3), 258-263 (1992).</li><li>Laughton, A. M., Garcia, J. R., Altincicek, B., Strand, M. R., Gerardo, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterisation+of+immune+responses+in+the+pea+aphid,+Acyrthosiphon+pisum.">Characterisation of immune responses in the pea aphid, Acyrthosiphon pisum.</a> J insect physiol. 57 (6), 830-839 (2011).</li><li>Barreaux, A. M. G., Barreaux, P., Koella, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Overloading+the+immunity+of+the+mosquito+Anopheles+gambiae+with+multiple+immune+challenges.">Overloading the immunity of the mosquito Anopheles gambiae with multiple immune challenges.</a> Parasite Vector. 9 (1), 210 (2016).</li><li>Lazzaro, B. P., Rolff, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Danger,+Microbes,+and+Homeostasis.">Danger, Microbes, and Homeostasis.</a> Science. 332 (6025), 43-44 (2011).</li><li>Arrighi, R. B. G., Faye, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasmodium+falciparum+GPI+toxin:+a+common+foe+for+man+and+mosquito.">Plasmodium falciparum GPI toxin: a common foe for man and mosquito.</a> Acta trop. 114 (3), 162-165 (2010).</li><li>Michel, K., Kafatos, F. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mosquito+immunity+against+Plasmodium.">Mosquito immunity against Plasmodium.</a> Insect Biochem Molec. 35 (7), 677-689 (2005).</li><li>Osta, M. A., Christophides, G. K., Vlachou, D., Kafatos, F. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Innate+immunity+in+the+malaria+vector+Anopheles+gambiae:+comparative+and+functional+genomics.">Innate immunity in the malaria vector Anopheles gambiae: comparative and functional genomics.</a> J Exp Biol. 207 (15), 2551-2563 (2004).</li><li>Christophides, G. K., Vlachou, D., Kafatos, F. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+and+functional+genomics+of+the+innate+immune+system+in+the+malaria+vector+Anopheles+gambiae.">Comparative and functional genomics of the innate immune system in the malaria vector Anopheles gambiae.</a> Immunol Rev. 198 (1), 127-148 (2004).</li><li>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=Mosquito+RNAi+is+the+major+innate+immune+pathway+controlling+arbovirus+infection+and+transmission.">Mosquito RNAi is the major innate immune pathway controlling arbovirus infection and transmission.</a> Future microbiol. 6 (3), 265-277 (2011).</li><li>Fragkoudis, R., Attarzadeh-Yazdi, G., Nash, A. A., Fazakerley, J. K., Kohl, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advances+in+dissecting+mosquito+innate+immune+responses+to+arbovirus+infection.">Advances in dissecting mosquito innate immune responses to arbovirus infection.</a> J Gen Virol. , (2009).</li><li>Schwartz, A., Koella, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+cost+of+immunity+in+the+yellow+fever+mosquito,+Aedes+aegypti+depends+on+immune+activation.">The cost of immunity in the yellow fever mosquito, Aedes aegypti depends on immune activation.</a> J evol biol. 17 (4), 834-840 (2004).</li><li>Lambrechts, L., Vulule, J. M., Koella, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+correlation+between+melanization+and+antibaterial+immune+responses+in+a+natural+population+of+the+malaria+vector+Anopheles+gambiae.">Genetic correlation between melanization and antibaterial immune responses in a natural population of the malaria vector Anopheles gambiae.</a> Evolution. 58 (10), 2377 (2004).</li><li>Boete, C., Paul, R. E. L., Koella, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+and+indirect+immunosuppression+by+a+malaria+parasite+in+its+mosquito+vector.">Direct and indirect immunosuppression by a malaria parasite in its mosquito vector.</a> P Roy Soc B-Biol Sci. 271 (1548), 1611-1615 (2004).</li><li>Sacks, D., Sher, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evasion+of+innate+immunity+by+parasitic+protozoa.">Evasion of innate immunity by parasitic protozoa.</a> Nat immunol. 3 (11), 1041-1047 (2002).</li><li>Zambrano-Villa, S., Rosales-Borjas, D., Carrero, J. C., Ortiz-Ortiz, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+protozoan+parasites+evade+the+immune+response.">How protozoan parasites evade the immune response.</a> Trend Parasito. 18 (6), 272-278 (2002).</li><li>Damian, R. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parasite+immune+evasion+and+exploitation:+reflections+and+projections.">Parasite immune evasion and exploitation: reflections and projections.</a> Parasitology. 115, S169-S175 (1997).</li><li>Schmid-Hempel, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parasite+immune+evasion:+a+momentous+molecular+war.">Parasite immune evasion: a momentous molecular war.</a> Trend ecol evol. 23 (6), 318-326 (2008).</li><li>Schmid-Hempel, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immune+defence,+parasite+evasion+strategies+and+their+relevance+for+&amp;#34;macroscopic+phenomena&amp;#34;+such+as+virulence.">Immune defence, parasite evasion strategies and their relevance for &amp;#34;macroscopic phenomena&amp;#34; such as virulence.</a> P Roy Soc B-Biol Sci. 364 (1513), 85-98 (2009).</li><li>Stearns, S. C., Koella, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+evolution+of+phenotypic+plasticity+in+life+history+traits-+predictions+of+reaction+norms+for+age+and+size+at+maturity.">The evolution of phenotypic plasticity in life history traits- predictions of reaction norms for age and size at maturity.</a> Evolution. 40 (5), 893-913 (1986).</li><li>Stearns, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Life-history+tactics:+a+review+of+the+ideas.">Life-history tactics: a review of the ideas.</a> Q rev biol. 51 (1), 3-47 (1976).</li><li>Valtonen, T. M., Kleino, A., Ramet, M., Rantala, 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=Starvation+Reveals+Maintenance+Cost+of+Humoral+Immunity.">Starvation Reveals Maintenance Cost of Humoral Immunity.</a> Evol Biol. 37 (1), 49-57 (2010).</li><li>Sheldon, B. C., Verhulst, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ecological+immunology:+costly+parasite+defences+and+trade-offs+in+evolutionary+ecology.">Ecological immunology: costly parasite defences and trade-offs in evolutionary ecology.</a> Trend Ecol Evo. 11 (8), 317-321 (1996).</li><li>Graham, A. L., Allen, J. E., Read, 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=Evolutionary+causes+and+consequences+of+immunopathology.">Evolutionary causes and consequences of immunopathology.</a> Annu Rev Ecol Evol S. 36, 373-397 (2005).</li><li>Best, A., Long, G., White, A., Boots, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+implications+of+immunopathology+for+parasite+evolution.">The implications of immunopathology for parasite evolution.</a> P Roy Soc B-Biol Sci. 279 (1741), 3234-3240 (2012).</li><li>Cator, L. J., George, 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=34;Manipulation&amp;#34;+without+the+parasite:+altered+feeding+behaviour+of+mosquitoes+is+not+dependent+on+infection+with+malaria+parasites.">34;Manipulation&amp;#34; without the parasite: altered feeding behaviour of mosquitoes is not dependent on infection with malaria parasites.</a> P Roy Soc B-Biol Sci. 280 (1763), 20130711 (2013).</li><li>Voordouw, M. J., Lambrechts, L., Koella, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=No+maternal+effects+after+stimulation+of+the+melanization+response+in+the+yellow+fever+mosquito+Aedes+aegypti.">No maternal effects after stimulation of the melanization response in the yellow fever mosquito Aedes aegypti.</a> Oikos. 117 (8), 1269-1279 (2008).</li><li>Paskewitz, S., Riehle, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Response+of+Plasmodium+refractory+and+susceptible+strains+of+Anopheles+gambiae+to+inoculated+Sephadex+beads.">Response of Plasmodium refractory and susceptible strains of Anopheles gambiae to inoculated Sephadex beads.</a> Dev comp immunol. 18 (5), 369-375 (1994).</li></ol>

The authors declare that they have no competing financial interests.

这种注射技术是很有用的激发和研究蚊子的黑化反应。例如，在这里我们研究的免疫刺激负荷的效果。 在此过程中的关键步骤是正确接种蚊子。以飞行肌或蚊子本身的任何过多的损害可能会阻止喂养蚊子或解剖之前，可能杀死它。第二个关键步骤是保持蚊子冰足够长的时间敲出来没有杀害他们。一个小容器将缓解和加快这一进程，因为它能够与冰更密切的联系。最后，在剥?...

昆虫依靠免疫反应，以保护自己免受寄生虫和致病菌1 - 3，通过他们的角质层或他们的肠上皮细胞4突破口。在蚊子，这些反应是对细菌5，6病毒，线虫丝虫7和疟原虫1,8,9高效。在蚊子，一个关键的免疫应答是外来颗粒与黑色素10的封装- 12。 12 -此封装可以在肠或血淋巴循环系统10发生。此黑化反应是亲酚的结果级联10 - 12，并且它可以导致寄生虫死亡或它们的吞噬作用。在成蚊，其中血细胞的细胞的数量是有限的，黑化是体液应答，如针对疟原虫寄生虫或丝虫线虫7。在其他一些昆虫，它直接是围绕寄生虫聚集melanize他们7血细胞的细胞。此外，黑色素也像鸡蛋的生产和表皮伤口愈合7其他几个生理过程是必不可少的。 免疫反应的刺激作为一种工具来研究昆虫干扰的几个农业和公共健康模型系统13 - 18。它在使用按蚊冈比亚蚊子，在非洲疟疾的主要载体，研究宿主-寄生虫的相互作用14 - 16,19。这些技术是基于昆虫以检测与它们的模式识别受体（PRR）2寄生虫的能力。蚊子也可以检测其他分子与其生物学如病原体相关分子模式（PAMP）的干扰，或检测由于胶原蛋白和核酸的释放自己的受损细胞。蚊子免疫细胞S，从而作为血细胞用于检测20 - 23。主免疫信号途径是Imd信号，收费，JAK / STAT 24和核糖核酸干扰（RNAi）25,26。这两种收费和IMD的途径影响黑色素的反应，并与亲酚互动级联10 - 12。 用于刺激黑色素响应的标准工具是用小珠进入胸腔的血淋巴蚊子的接种。黑色素包封的程度可以然后通过蚊子的解剖检索胎圈后测量19。在大多数研究中，只有一个小珠被每蚊15,16,27注入，但注入多珠是可能的，以研究在黑化响应19的限制。这些珠粒使用注射液（生理血清）来限制蚊子生理学紊乱注入和蚊子15,16,27的干燥。的染料加入到该溶液中，以缓解珠选择。它是用于检索珠子15,16,27解剖溶液是相同的。 用非致病性刺激接种昆虫的优点是把重点放在对免疫应答的直接作用的能力。有因寄生虫致病28，免疫抑制29没有复杂影响- 31，或免疫逃避31 - 34。此外，与其他生活史性状，如长寿或繁殖力的刺激所造成的后果，也可以研究。因此，研究人员在研究进化生态学可能需要这样的工具2,35,36。例如，免疫挑战黄蜂有饥饿状态下寿命缩短。免疫刺激和部署类似的负面影响已经在不同的无脊椎动物模型中观察到，往往导致在很短的呃寿命或更少的繁殖成功13,27,37。这些研究可以在不同的环境中2,4,38进行。刺激免疫力也感兴趣的那些直接专注于免疫病理学39,40。 这个协议是基于珠子的接种的蚊子刺激黑化反应，并直接测量黑色素的量。这使得在不同的实验设置黑化反应的定量和定性研究。这样的工具可以扩展到其它的免疫反应，如抗菌响应于刺激热灭活细菌41。它也可以在许多生态设置进行。

<table><tbody><tr><td>Microcapillary glass tubes GB120TF-10</td><td>science-products.com</td><td>GB120TF-10</td><td>http://www.science-products.com/Products/CatalogG/Glass/Glass.html</td></tr><tr><td>Microcaps Capillary pipette bulb</td><td>Drumond</td><td>1-000-9000</td><td></td></tr><tr><td>negatively charged Sephadex CM C-25 beads</td><td>Sigma-Aldrich, Steinheim, Germany</td><td>C25120 SIGMA</td><td>need few to start</td></tr><tr><td>Methyl green</td><td>Sigma-Aldrich</td><td>323829 ALDRICH</td><td>need few to start</td></tr><tr><td>Software ImageJ</td><td>opensource</td><td></td><td>Version 1.47f7 or later</td></tr></tbody></table>

inoculating anopheles gambiae mosquitoes with beads to induce and measure the melanization immune response

免疫反应的刺激是无脊椎动物研究的常用工具来检查的有效性和免疫力的机制。这种刺激是基于注射非致病性颗粒的成虫，如颗粒将被免疫系统检测，将诱导产生免疫效应的。在这里，我们专注于在蚊子冈比亚按蚊黑化反应的刺激。黑化反应的结果在外来颗粒和寄生虫与黑色素的暗层封装。为了刺激这个回应，蚊子接种使用微细玻璃管在胸腔珠。然后，24小时后，蚊子解剖检索珠。胎圈的黑化的程度是使用图像分析软件来测量。珠没有寄生虫的致病作用，或它们的容量来逃避或抑制免疫应答。这些注射是measu方式再免疫效果和免疫刺激对其他生活史特征，如孕卵或寿命的影响。这是不完全一样的直接研究宿主 - 寄生虫的相互作用，但它是一个有趣的工具来研究免疫和其进化生态学。

<html>蚊子没有所有melanize以同样的方式的珠，如一些珠粒少覆盖有黑色素比其他（ 图1）。事实上，一些珠子由于缺乏黑色素的仍然蓝色，但其他人却完全黑了（ 图1）。黑化值通过线性内插标准化为值0（这对应于蓝色和unmelanized珠）和100（对应于一个黑暗和严重白化珠）（ 图2a和2b）之间。这些值表示的珠粒黑色素的程度...

Watch this Scientific Journal Video about Inoculating Anopheles gambiae Mosquitoes with Beads to Induce and Measure the Melanization Immune Response at JoVE.com

Inoculating Anopheles gambiae Mosquitoes with Beads to Induce and Measure the Melanization Immune Response

Through inoculation with beads, the described technique enables the stimulation of the mosquito melanization response in the hemolymph circulating system. The amount of melanin covering the beads can be measured after dissection as a measure of the immune response.

接种<em&gt;冈比亚按蚊</em&gt;珠蚊子诱发和测量黑化免疫反应

The stimulation of immune responses is a common tool in invertebrate studies to examine the efficacy and the mechanisms of immunity. This stimulation is ...

inoculating-anopheles-gambiae-mosquitoes-with-beads-to-induce-measure

Research

JoVE Journal

Immunology and Infection

Inoculating Anopheles gambiae Mosquitoes with Beads to Induce and Measure the Melanization Immune Response

7.9K 次观看.  Université de Neuchâtel. Through inoculation with beads, the described technique enables the stimulation of the mosquito melanization response in the hemolymph circulating system. The amount of melanin covering the beads can be measured after dissection as a measure of the immune response.

视频: Inoculating Anopheles gambiae Mosquitoes with Beads to Induce and Measure the Melanization Immune Response

Injection of An. stephensi Embryos to Generate Malaria-resistant Mosquitoes

The introduction of exogenous genes into the genomes of mosquitoes requires microinjection techniques tailored to the specific species of interest. This video protocol demonstrates a method used by the James laboratory to microinject DNA constructs into Anopheles stephensi embryos for the generation of transformed mosquitoes. Techniques for preparing microinjection needles, collecting and preparing embryos and performing the microinjection are illustrated.

Anopheles stephensi mosquitoes are vectors for malaria inhabiting India and throughout Asia. This video demonstrates the technique for performing microinjections of this species with transgenes that will confer resistance to the malaria to the mosquito. Much of the methodology demonstrated in this video is applicable to microinjection techniques of other mosquito species.

注射的。 stephensi胚胎生成抗药性疟疾的蚊子

The introduction of exogenous genes into the genomes of mosquitoes requires microinjection techniques tailored to the specific species of interest. This ...

科研

生物学

Site-Directed &#966;C31-Mediated Integration and Cassette Exchange in Anopheles Vectors of Malaria

Functional genomic analysis and related strategies for genetic control of malaria rely on validated and reproducible methods to accurately modify the genome of Anopheles mosquitoes. Amongst these methods, the &#966;C31 system allows precise and stable site-directed integration of transgenes, or the substitution of integrated transgenic cassettes via recombinase-mediated cassette exchange (RMCE). This method relies on the action of the Streptomyces &#966;C31 bacteriophage integrase to catalyze recombination between two specific attachment sites designated attP (derived from the phage) and attB (derived from the host bacterium). The system uses one or two attP sites that have been integrated previously into the mosquito genome and attB site(s) in the donor template DNA. Here we illustrate how to stably modify the genome of attP-bearing Anopheles docking lines using two plasmids: an attB-tagged donor carrying the integration or exchange template and a helper plasmid encoding the &#966;C31 integrase. We report two representative results of &#966;C31-mediated site-directed modification: the single integration of a transgenic cassette in An. stephensi and RMCE in An. gambiae mosquitoes. &#966;C31-mediated genome manipulation offers the advantage of reproducible transgene expression from validated, fitness neutral genomic sites, allowing comparative qualitative and quantitative analyses of phenotypes. The site-directed nature of the integration also substantially simplifies the validation of the single insertion site and the mating scheme to obtain a stable transgenic line. These and other characteristics make the &#966;C31 system an essential component of the genetic toolkit for the transgenic manipulation of malaria mosquitoes and other insect vectors.

The protocol describes how to achieve site-directed modifications in the genome of Anopheles malaria mosquitoes using the &#966;C31 system. Modifications described include both the integration and the exchange of transgenic cassettes in the genome of attP-bearing docking lines.

位点导向 的φC31介导的疟疾 按蚊 载体中的整合和盒式交换

Functional genomic analysis and related strategies for genetic control of malaria rely on validated and reproducible methods to accurately modify the ...

遗传学

Microinjection Method for Anopheles gambiae Embryos

Embryo microinjection techniques are essential for many molecular and genetic studies of insect species. They provide a means to introduce exogenous DNA fragments encoding genes of interest as well as favorable traits into the insect germline in a stable and heritable manner. The resulting transgenic strains can be studied for phenotypic changes resulting from the expression of the integrated DNA to answer basic questions or used in practical applications. Although the technology is straightforward, it requires of the investigator patience and practice to achieve a level of skill that maximizes efficiency. Shown here is a method for microinjection of embryos of the African malaria mosquito, Anopheles gambiae. The objective is to deliver by microinjection exogenous DNA to the embryo so that it can be taken up in the developing germline (pole) cells. Expression from the injected DNA of transposases, integrases, recombinases, or other nucleases (for example CRISPR-associated proteins, Cas) can trigger events that lead to its covalent insertion into chromosomes. Transgenic An. gambiae generated from these technologies have been used for basic studies of immune system components, genes involved in blood-feeding, and elements of the olfactory system. In addition, these techniques have been used to produce An. gambiae strains with traits that may help control the transmission of malaria parasites.

Microinjection Method for Anopheles gambiae Embryos

Microinjection techniques are essential to introduce exogenous genes into the genomes of mosquitoes. This protocol explains a method used by the James laboratory to microinject DNA constructs into Anopheles gambiae embryos to generate transformed mosquitoes.

冈 比亚按蚊 胚胎的显微注射方法

Embryo microinjection techniques are essential for many molecular and genetic studies of insect species. They provide a means to introduce exogenous DNA ...

An In Vitro Model for Measuring Immune Responses to Malaria in the Context of HIV Co-infection

Malaria and HIV co-infection is a growing health priority. However, most research on malaria or HIV currently focuses on each infection individually. Although understanding the disease dynamics for each of these pathogens independently is vital, it is also important that the interactions between these pathogens are investigated and understood. 
We have developed a versatile in vitro model of HIV-malaria co-infection to study host immune responses to malaria in the context of HIV infection. Our model allows the study of secreted factors in cellular supernatants, cell surface and intracellular protein markers, as well as RNA expression levels. The experimental design and methods used limit variability and promote data reliability and reproducibility. 
All pathogens used in this model are natural human pathogens (Plasmodium falciparum and HIV-1), and all infected cells are naturally infected and used fresh. We use human erythrocytes parasitized with P. falciparum and maintained in continuous in vitro culture. We obtain freshly isolated peripheral blood mononuclear cells from chronically HIV-infected volunteers. Every condition used has an appropriate control (P. falciparum parasitized vs. normal erythrocytes), and every HIV-infected donor has an HIV uninfected control, from which cells are harvested on the same day. This model provides a realistic environment to study the interactions between malaria parasites and human immune cells in the context of HIV infection.

An In Vitro Model for Measuring Immune Responses to Malaria in the Context of HIV Co-infection

Human co-infection is difficult to replicate in vitro. However, human malaria parasites can readily be cultured in vitro, as can freshly isolated human peripheral blood mononuclear cells naturally infected with HIV. This provides an excellent model for studying early immune responses to malaria parasites in the context of HIV co-infection.

一个<em&gt;体外</em&gt;模型在HIV合并感染的背景下测量免疫应答疟疾

Malaria and HIV co-infection is a growing health priority. However, most research on malaria or HIV currently focuses on each infection individually. ...

免疫与感染

Myeloid Cell Isolation from Mouse Skin and Draining Lymph Node Following Intradermal Immunization with Live Attenuated Plasmodium Sporozoites

Malaria infection begins when the sporozoite stage of Plasmodium is inoculated into the skin of a mammalian host through a mosquito bite. The highly motile parasite not only reaches the liver to invade hepatocytes and transform into erythrocyte-infective form. It also migrates into the skin and to the proximal lymph node draining the injection site, where it can be recognized and degraded by resident and/or recruited myeloid cells. Intravital imaging reported the early recruitment of brightly fluorescent Lys-GFP positive leukocytes in the skin and the interactions between sporozoites and CD11c+ cells in the draining lymph node. We present here an efficient procedure to recover, identify and enumerate the myeloid cell subsets that are recruited to the mouse skin and draining lymph node following intradermal injection of immunizing doses of sporozoites in a murine model. Phenotypic characterization using multi-parametric flow cytometry provides a reliable assay to assess early dynamic cellular changes during inflammatory response to Plasmodium infection.

Myeloid Cell Isolation from Mouse Skin and Draining Lymph Node Following Intradermal Immunization with Live Attenuated Plasmodium Sporozoites

We describe here a protocol for isolating myeloid cells from mouse skin and draining lymph node following intradermal injection of Plasmodium sporozoites. Flow cytometry of collected cells provides a reliable assay to characterize the skin and draining lymph node inflammatory response to the parasite.

从老鼠皮肤及引流淋巴结髓细胞分离皮下继免疫减毒活<em&gt;疟原虫</em&gt;孢子

Malaria infection begins when the sporozoite stage of Plasmodium is inoculated into the skin of a mammalian host through a mosquito bite. The highly ...

In Vivo Tracking of Edema Development and Microvascular Pathology in a Model of Experimental Cerebral Malaria Using Magnetic Resonance Imaging

Cerebral malaria is a sign of severe malarial disease and is often a harbinger of death. While aggressive management can be life-saving, the detection of cerebral malaria can be difficult. We present an experimental mouse model of cerebral malaria that shares multiple features of the human disease, including edema and microvascular pathology. Using magnetic resonance imaging (MRI), we can detect and track the blood-brain barrier disruption, edema development, and subsequent brain swelling. We describe multiple MRI techniques that can visualize these pertinent pathological changes. Thus, we show that MRI represents a valuable tool to visualize and track pathological changes, such as edema, brain swelling, and microvascular pathology, in vivo.

In Vivo Tracking of Edema Development and Microvascular Pathology in a Model of Experimental Cerebral Malaria Using Magnetic Resonance Imaging

We describe a mouse model of experimental cerebral malaria and show how inflammatory and microvascular pathology can be tracked in vivo using magnetic resonance imaging.

使用磁共振成像实验性脑疟疾模型中的水肿发育​​和微血管病理学的体内追踪

Cerebral malaria is a sign of severe malarial disease and is often a harbinger of death. While aggressive management can be life-saving, the detection of ...

医学

Standard Membrane Feeding Assay for the Detection of Plasmodium falciparum Infection in Anopheles Mosquito Vectors

Malaria remains one of the most devastating diseases worldwide and, to date, the African region is still responsible for 94% of all cases worldwide. This parasitic disease requires a protozoan parasite, an Anopheles mosquito vector, and a vertebrate host. The Anopheles genus comprises more than 500 species, of which 60 are known as vectors of the parasite. The Plasmodium parasite genus consists of 250 species, and 48 of these are involved in disease transmission. Furthermore, the Plasmodium falciparum parasite has contributed toward an estimated 99.7% of malaria cases in sub-Saharan Africa in recent years. 
Gametocytes form part of the sexual stage of the parasite and are ingested by the female mosquito upon feeding on an infected human host. Further development of the parasite within the mosquito is enhanced by favorable environmental conditions in the midgut of the mosquito. Here, the fusion of the female and male gametes takes place, and the motile ookinetes originate. The ookinetes enter the midgut epithelium of the mosquito, and mature ookinetes form oocysts, which, in turn, produce motile sporozoites. These sporozoites migrate to the mosquito's salivary glands and are injected as a mosquito takes a blood meal. 
For drug discovery purposes, mosquitoes were artificially infected with gametocyte-infected blood in the standard membrane feeding assay (SMFA). To detect infection within the mosquito and/or to assess the efficacy of antimalarial compounds, the midguts of the female mosquitoes were removed post infection and were stained with mercurochrome. This method was used to enhance the visual detection of oocysts under the microscope for the accurate determination of oocyst prevalence and intensity.

Standard Membrane Feeding Assay for the Detection of Plasmodium falciparum Infection in Anopheles Mosquito Vectors

The standard membrane feeding assay (SMFA) is regarded as the gold standard for the assessment and identification of potential antimalarial compounds. This artificial feeding system is used to infect mosquitoes to further evaluate the effects of such compounds on the intensity and prevalence of the Plasmodium falciparum parasite.

用于检测按蚊媒介中恶性疟原虫感染的标准膜喂养测定

Malaria remains one of the most devastating diseases worldwide and, to date, the African region is still responsible for 94% of all cases worldwide. This ...

Generating Genetically Modified Plasmodium berghei Sporozoites

Malaria is a deadly disease caused by the parasite Plasmodium and is transmitted through the bite of female Anopheles mosquitoes. The sporozoite stage of Plasmodium deposited by mosquitoes in the skin of vertebrate hosts undergoes a phase of mandatory development in the liver before initiating clinical malaria. We know little about the biology of Plasmodium development in the liver; access to the sporozoite stage and the ability to genetically modify such sporozoites are critical tools for studying the nature of Plasmodium infection and the resulting immune response in the liver. Here, we present a comprehensive protocol for the generation of transgenic Plasmodium berghei sporozoites. We genetically modify blood-stage P. berghei and use this form to infect Anopheles mosquitoes when they take a blood meal. After the transgenic parasites undergo development in the mosquitoes, we isolate the sporozoite stage of the parasite from the mosquito salivary glands for in vivo and in vitro experimentation. We demonstrate the validity of the protocol by generating sporozoites of a novel strain of P. berghei expressing the green fluorescent protein (GFP) subunit 11 (GFP11), and show how it could be used to investigate the biology of liver-stage malaria.

Generating Genetically Modified Plasmodium berghei Sporozoites

Malaria is transmitted through inoculation of the sporozoite stage of Plasmodium by infected mosquitoes. Transgenic Plasmodium has allowed us to understand the biology of malaria better and has contributed directly to malaria vaccine development efforts. Here, we describe a streamlined methodology to generate transgenic Plasmodium berghei sporozoites.

生成转基因 伯氏疟原 虫孢子

Malaria is a deadly disease caused by the parasite Plasmodium and is transmitted through the bite of female Anopheles mosquitoes. The sporozoite stage of ...

RNAi Trigger Delivery into Anopheles gambiae Pupae

RNA interference (RNAi), a naturally occurring phenomenon in eukaryotic organisms, is an extremely valuable tool that can be utilized in the laboratory for functional genomic studies. The ability to knockdown individual genes selectively via this reverse genetic technique has allowed many researchers to rapidly uncover the biological roles of numerous genes within many organisms, by evaluation of loss-of-function phenotypes. In the major human malaria vector Anopheles gambiae, the predominant method used to reduce the function of targeted genes involves injection of double-stranded (dsRNA) into the hemocoel of the adult mosquito. While this method has been successful, gene knockdown in adults excludes the functional assessment of genes that are expressed and potentially play roles during pre-adult stages, as well as genes that are expressed in limited numbers of cells in adult mosquitoes. We describe a method for the injection of Serine Protease Inhibitor 2 (SRPN2) dsRNA during the early pupal stage and validate SRPN2 protein knockdown by observing decreased target protein levels and the formation of melanotic pseudo-tumors in SRPN2 knockdown adult mosquitoes. This evident phenotype has been described previously for adult stage knockdown of SRPN2 function, and we have recapitulated this adult phenotype by SRPN2 knockdown initiated during pupal development. When used in conjunction with a dye-labeled dsRNA solution, this technique enables easy visualization by simple light microscopy of injection quality and distribution of dsRNA in the hemocoel.

RNAi Trigger Delivery into Anopheles gambiae Pupae

RNA interference (RNAi) is an extremely valuable tool for uncovering gene function. However, the ability to target genes using RNAi during pre-adult stages is limited in the major human malaria vector Anopheles gambiae. We describe an RNAi protocol to reduce gene function via direct injection during pupal development.

RNAi的触发传送到<em&gt;冈比亚按蚊</em&gt;蛹

RNA interference (RNAi), a naturally occurring phenomenon in eukaryotic organisms, is an extremely valuable tool that can be utilized in the laboratory ...

Effective Oral RNA Interference (RNAi) Administration to Adult Anopheles gambiae Mosquitoes

RNA interference has been a heavily utilized tool for reverse genetic analysis for two decades. In adult mosquitoes, double-stranded RNA (dsRNA) administration has been accomplished primarily via injection, which requires significant time and is not suitable for field applications. To overcome these limitations, here we present a more efficient method for robust activation of RNAi by oral delivery of dsRNA to adult Anopheles gambiae. Long dsRNAs were produced in Escherichia coli strain HT115 (DE3), and a concentrated suspension of heat-killed dsRNA-containing bacteria in 10% sucrose was offered on cotton balls ad-libitum to adult mosquitoes. Cotton balls were replaced every 2 days for the duration of the treatment. Use of this method to target doublesex (a gene involved in sex differentiation) or fork head (which encodes a salivary gland transcription factor) resulted in reduced target gene expression and/or protein immunofluorescence signal, as measured by quantitative Real-Time PCR (qRT-PCR) or fluorescence confocal microscopy, respectively. Defects in salivary gland morphology were also observed. This highly flexible, user-friendly, low-cost, time-efficient method of dsRNA delivery could be broadly applicable to target genes important for insect vector physiology and beyond.

Effective Oral RNA Interference RNAi Administration to Adult Anopheles gambiae Mosquitoes

The oral administration of dsRNA produced by bacteria, a delivery method for RNA interference (RNAi) that is routinely used in Caenorhabditis elegans, was successfully applied here to adult mosquitoes. Our method allows for robust reverse genetics studies and transmission-blocking vector studies without the use of injection.

接种

摘要

探索更多视频

此视频中的章节

注射的。 stephensi胚胎生成抗药性疟疾的蚊子

位点导向的φC31介导的疟疾按蚊载体中的整合和盒式交换

冈比亚按蚊胚胎的显微注射方法

一个

从老鼠皮肤及引流淋巴结髓细胞分离皮下继免疫减毒活

使用磁共振成像实验性脑疟疾模型中的水肿发育和微血管病理学的体内追踪

用于检测按蚊媒介中恶性疟原虫感染的标准膜喂养测定

生成转基因伯氏疟原虫孢子

RNAi的触发传送到

对成年冈比亚按蚊的有效口服RNA干扰（RNAi）给药

接种

摘要

探索更多视频

此视频中的章节

注射的。 stephensi胚胎生成抗药性疟疾的蚊子

位点导向 的φC31介导的疟疾 按蚊 载体中的整合和盒式交换

冈 比亚按蚊 胚胎的显微注射方法

一个

从老鼠皮肤及引流淋巴结髓细胞分离皮下继免疫减毒活

使用磁共振成像实验性脑疟疾模型中的水肿发育​​和微血管病理学的体内追踪

用于检测按蚊媒介中恶性疟原虫感染的标准膜喂养测定

生成转基因 伯氏疟原 虫孢子

RNAi的触发传送到

对成年 冈比亚按蚊 的有效口服RNA干扰（RNAi）给药

位点导向的φC31介导的疟疾按蚊载体中的整合和盒式交换

冈比亚按蚊胚胎的显微注射方法

使用磁共振成像实验性脑疟疾模型中的水肿发育和微血管病理学的体内追踪

生成转基因伯氏疟原虫孢子

对成年冈比亚按蚊的有效口服RNA干扰（RNAi）给药