Name: extraction of hemocytes from drosophila melanogaster larvae for microbial infection and analysis
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MCherry ifade Coxiella burnetiistokları sağlamak için Dr Robert Heinzen için minnettarız. Biz Dr Luis Teixeira omurgasız yanardöner virüs 6 ve Bloomington stok merkezi sinek stokları sağlamak için verdiğiniz için teşekkür ederiz. Bu projenin kısmen NIH hibe R00 AI106963 (A.G.G.) ve Washington State Üniversitesi tarafından finanse edildi.

1. ex vivo enfeksiyon
<ol><li>Orta ve donanımları<ol>
<li>Steril koşullarda, taze Drosophila Hemocyte izole Orta (% 75 Schneider'ın Drosophila içeren DHIM) hazırlamak orta % 25 ile Fetal sığır Serum (FBS) ve filtre bu sterilize.</li>
		<li>2-3 adet 10 cm x 10 cm parafin film bir stereomicroscope altında katman.</li>
		<li>Cam kılcal hazırlayın. Kapiller çektirme ısıtıcı maksimum % 55'i için ayarlayın. Kılcal tüp yaklaşık 10 µm keskin bir noktaya kadar çekin.</li>
		<...

<ol>
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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+hemocyte+load+is+associated+with+increased+resistance+against+parasitoids+in+Drosophila+suzukii,+a+relative+of+D.+melanogaster.">High hemocyte load is associated with increased resistance against parasitoids in Drosophila suzukii, a relative of D. melanogaster.</a> PLoS One. 7 (4), 34721 (2012).</li><li>Tsuzuki, S., 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=Switching+between+humoral+and+cellular+immune+responses+in+Drosophila.+is+guided+by+the+cytokine+GBP.">Switching between humoral and cellular immune responses in Drosophila. is guided by the cytokine GBP.</a> Nat Commun. 5, 4628 (2014).</li><li>Kurucz, 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=Definition+of+Drosophila.+hemocyte+subsets+by+cell-type+specific+antigens.">Definition of Drosophila. hemocyte subsets by cell-type specific antigens.</a> Acta Biol Hung. 58, 95-111 (2007).</li><li>Neyen, C., Bretscher, A. 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Immune System and Leads to a Pro-Inflammatory State.</a> PLoS One. 10 (8), 0136593 (2015).</li><li>Rus, F., 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=Expression+pattern+of+Filamin-240+in+Drosophila+blood+cells.">Expression pattern of Filamin-240 in Drosophila blood cells.</a> Gene Expr Patterns. 6 (8), 928-934 (2006).</li><li>Kurucz, 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=Hemese,+a+hemocyte-specific+transmembrane+protein,+affects+the+cellular+immune+response+in+Drosophila.">Hemese, a hemocyte-specific transmembrane protein, affects the cellular immune response in Drosophila.</a> P Natl Acad Sci USA. 100 (5), 2622-2627 (2003).</li><li>Markus, 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=Sessile+hemocytes+as+a+hematopoietic+compartment+in+Drosophila+melanogaster.">Sessile hemocytes as a hematopoietic compartment in Drosophila melanogaster.</a> P Natl Acad Sci USA. 106 (12), 4805-4809 (2009).</li><li>Bretscher, A. 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Nasıl konak hücreleri enfekte olma daha iyi anlamak için özellikle daha önce denenmemiş patojen ve hücre tipi kombinasyonları4' te deneme zaman patojen hücrelerdeki lokalizasyonu netleştirmek önemlidir. Enfeksiyonu takip hücresel yanıt basamaklı eğitim üretken patojen işgali gösterebilir iken, görüntüleme ve hücresel yanıt verilerini kombinasyonu patojen işgali ve enfeksiyon göstermek için önemlidir. Patojen işgalinin 2D görüntüleri ana hücreleri gösteren raporlar ...

Drosophila melanogaster doğuştan gelen bağışıklık1incelenmesi için bir iyi kurulmuş model organizmadır. Doğuştan gelen bağışıklık yanıtı sırasında hemocytes yanıt patojeni meydan okuma olarak önemli bir rol oynamaktadır. Hemocytes mantar, viral ve bakteriyel enfeksiyon2,3sırasında fagositik eylem yoluyla patojen mücadelede önemli bir işleve sahip yanı sıra parazit Kapsüllenen için kritik öneme sahiptir.
En iyi ana bilgisayarın doğuştan gelen bağışıklık yanıtı patojenik mikrobiyal enfeksiyon anlamak için nasıl patojen konak hücreleri enfeksiyon sırasında işgal görselleştirmek önemlidir. Bu görselleştirme işgali mekanizmasının bir anlaşmaya katkıda bulunur. Patojen hücre içi yerelleştirme ayrıntılarını ve hücresel yanıt ile birlikte, bu veriler enfeksiyon ve hangi ile mikrop etkileşim hücre organelleri ana bilgisayar yanıt hakkında ipuçları sağlar. Böylece, 3D modeli yeniden görüntüleme mikroskobu tarafından sonra konak hücreleri patojenler tam yerini belirlemek yararlı olabilir. Bu çalışmada, Avusturya'nın Coxiella burnetii (C. burnetii), Q ateşi, insan ve hayvan sağlığı için ciddi bir tehdit teşkil etmektedir birincil Drosophila hemocytes Hayvansal bir hastalık hastalığının görüntülenir. Son zamanlarda, Drosophila 2 dokuz mil faz II (NMII) klon C. burnetii 4 suşu ve bu zorlanma Drosophila4' te, çoğaltmak yapabiliyor bu gösteren Biyogüvenlik düzeyi duyarlı gösterilmiştir Drosophila C. burnetii patogenezi okumaya bir model organizma kullanılır.
Önceki çalışmalarda hemocytes ana bilgisayarın doğuştan gelen bağışıklık yanıtı incelemek için kullandık. Hemocytes kullanılan morfolojik gözlemleri5,6,7, xarakteristikaları analiz2,8, fagositoz analiz2,3, qRT-PCR2 için , 9, immunoprecipitation10,11, immünfloresan analiz10,12, immunostaining13, immunoblotting3,10, 11 ve immünhistokimya9,14. Drosophila S2 hücreleri de çeşitli vitro deneyler için kullanılabilir olmakla birlikte, onların davranış15,16-əbadoləşdirmək ve önceden var olan potansiyel viral enfeksiyon değiştirmek. Primer Hücre S2 hücreleri gibi bir ölümsüzleştirdi hücre kültürünü aksine kullanımı doğuştan gelen bağışıklık fonksiyonu çalışma için bir sistem tüm organizmanın daha temsilcisi sağlar. Ayrıca, hemocytes vivo, ayıklama önce enfeksiyonu hücrelere diğer ana bilgisayar proteinler ve doku, hemocytes ex vivo enfeksiyonu önce çıkarılması üzerinde bir avantaj etkileşim sağlar. Birkaç farklı yöntem hemocytes yeterli sayıda hemocytes canlı8,17,18,19tutmak için zaman kısa bir süre içinde elde etmek için kullanılmıştır.
Bu çalışmada, biz Drosophila 3rd INSTAR larva C. burnetii, Listeria Monositogenez (Listeria) veya omurgasız yanardöner patojenik mikrobiyal enfeksiyon için hemocytes ayıklamak için bir yöntem mevcut virüs 6 (IIV6). Vivo ve ex vivo hemocyte enfeksiyon yöntemleri açıklanmaktadır. Vivo- ve ex vivo-virüslü hemocytes confocal mikroskobu ile görüntülenir ve C. burnetii işgalinin 3D modeller oluşturmak için kullanılır. Buna ek olarak, ex vivoayıklama iletişim kuralını kullanarak-virüslü hemocytes için gen ve protein ifadesi kullanılmıştır deneyleri. Özellikle, IIV6 ve Listeriaenfeksiyonu ölçüde incelemek için toplam RNA veya protein qRT-PCR veya Western blot analizi için hücrelerden izole edildi. Birlikte ele alındığında, protokol hızla hemocytes sayısının fazlalığı 3rd INSTAR larva ve birincil hemocytes, in vivo veya ex vivoenfekte kanıt toplamak için yöntemler sağlar, için uygun bir platform olan mikrobiyal patojen enfeksiyon çalışmaları ve ilgili aşağı akım analizleri mikroskobu, transcriptomics ve proteomik gibi.

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	<tbody>
		<tr height="43">
			<td height="43" style="height:43px;width:303px;">Schneider&#39;s Drosophila Medium&#160;</td>
			<td style="width:288px;">Thermo Fisher Scientific (Gibco)</td>
			<td style="width:145px;">21720024</td>
			<td style="width:179px;">1.1.1), 2.1.2)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Fetal Bovine Serum</td>
			<td style="width:288px;">GE Healthcare Life Sciences&#160; (HyClone)</td>
			<td style="width:145px;">SH30070.03HI</td>
			<td style="width:179px;">1.1.1), 2.1.2)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Filter (0.22 &#181;L)</td>
			<td style="width:288px;">RESTEK</td>
			<td style="width:145px;">26158</td>
			<td style="width:179px;">1.1.1)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Strainer (100 &#181;m)</td>
			<td style="width:288px;">Greiner bio-one</td>
			<td style="width:145px;">542000</td>
			<td>1.2.1), 2)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Stereo microscope</td>
			<td style="width:288px;">Amscope</td>
			<td style="width:145px;">SM-1BSZ-L6W</td>
			<td style="width:179px;">1.2), 2)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Glass capillary</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:145px;">21-171-4</td>
			<td style="width:179px;">1.1), 1.2), 2)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:303px;">Capillary puller</td>
			<td style="width:288px;">Narishige International USA, Inc.</td>
			<td style="width:145px;">PC-10</td>
			<td style="width:179px;">1.1.3)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Mineral oil</td>
			<td style="width:288px;">Snow River Products</td>
			<td style="width:145px;"></td>
			<td style="width:179px;">1.1.4)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Nanoinjector</td>
			<td style="width:288px;">Drummond Scientific Company</td>
			<td style="width:145px;">3-000-204</td>
			<td style="width:179px;">1.1), 1.2), 2.2)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Forceps</td>
			<td style="width:288px;">VWR</td>
			<td style="width:145px;">82027-402</td>
			<td style="width:179px;">1.1.5), 1.2), 2), 3.1.7)</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:303px;">CO2 delivery apparatus</td>
			<td style="width:288px;">Genesee Scientific</td>
			<td style="width:145px;">59-122BC</td>
			<td style="width:179px;">1.2), 2)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:303px;">Trypan Blue</td>
			<td style="width:288px;">Thermo Fisher Scientific (Gibco)</td>
			<td style="width:145px;">15250061</td>
			<td style="width:179px;">1.3)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Hemocytometer</td>
			<td style="width:288px;">Hausser Scientific</td>
			<td style="width:145px;">3100</td>
			<td style="width:179px;">1.3)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">24 well plate</td>
			<td style="width:288px;">Greiner bio-one</td>
			<td style="width:145px;">662160</td>
			<td style="width:179px;">1.4), 2.2)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Coxiella burnetii - mCherry</td>
			<td style="width:288px;">Dr. Heinzen, R.</td>
			<td style="width:145px;"></td>
			<td style="width:179px;">1.4), 2.2)</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:303px;">Drosophila fruit juice plates</td>
			<td style="width:288px;">Cold Spring Harbor Protocols</td>
			<td style="width:145px;"></td>
			<td>2.1) http://cshprotocols.cshlp.org/content/2007/9/pdb.rec11113.full</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:303px;">Agar</td>
			<td style="width:288px;">Fisher Bioreagents</td>
			<td style="width:145px;">BP1423-500</td>
			<td style="width:179px;">2.1.1.1)</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:303px;">Methyl paraben</td>
			<td style="width:288px;">Amresco</td>
			<td style="width:145px;">0572-500G</td>
			<td style="width:179px;">2.1.1.2)</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:303px;">Absolute ethanol</td>
			<td style="width:288px;">Fisher Bioreagents</td>
			<td style="width:145px;">BP2818-500</td>
			<td style="width:179px;">2.1.1.2)</td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:303px;">Welch&#39;s 100% Grape juice frozen concentrate, 340 mL</td>
			<td style="width:288px;">Amazon</td>
			<td style="width:145px;">B0025UJVGM</td>
			<td style="width:179px;">2.1.1.3)</td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:303px;">Petri dishes, 10 x 35 mm</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:145px;">08-757-100A</td>
			<td style="width:179px;">2.1.1.4)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Microscope cover glass</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:145px;">12-545-80</td>
			<td style="width:179px;">1.4.4), 2.2.2)</td>
		</tr>
		<tr height="70">
			<td height="70" style="height:70px;width:303px;">Yeast, Bakers Dried Active</td>
			<td style="width:288px;">MP Biomedicals</td>
			<td style="width:145px;">0210140001</td>
			<td>2.1) Add 2 parts of water to 1 part of yeast (v/v)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Tungsten needle</td>
			<td style="width:288px;">Fine Science Tools</td>
			<td style="width:145px;">10130-20</td>
			<td style="width:179px;">2.1)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Holding forceps</td>
			<td style="width:288px;">VWR</td>
			<td style="width:145px;">HS8313</td>
			<td style="width:179px;">2.1)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Paraformaldehyde</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:145px;">FLO4042-500</td>
			<td style="width:179px;">3.1.3)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Triton X-100</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:145px;">BP151-500</td>
			<td style="width:179px;">3.1.3)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Bovine Serum Albumin</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:145px;">BP9706-100</td>
			<td style="width:179px;">3.1.3)</td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:303px;">4&#39;,6-diamidino-2-phenylindole</td>
			<td style="width:288px;">Thermo Fisher Scientific</td>
			<td style="width:145px;">62247</td>
			<td style="width:179px;">3.1.4)</td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:303px;">Antifade mounting medium</td>
			<td style="width:288px;">Thermo Fisher Scientific</td>
			<td style="width:145px;">P36930</td>
			<td style="width:179px;">3.1.6)</td>
		</tr>
		<tr height="132">
			<td height="132" style="height:132px;width:303px;">Confocal microsope</td>
			<td style="width:288px;">Leica</td>
			<td style="width:145px;">TCS SP8-X White Light Confocal Laser Scanning Microscope</td>
			<td style="width:179px;">3.2)</td>
		</tr>
		<tr height="103">
			<td height="103" style="height:103px;width:303px;">3D imaging reconstruction software</td>
			<td style="width:288px;">Leica</td>
			<td style="width:145px;">LASX with 3D visualization module</td>
			<td style="width:179px;">3.3)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Microscope slides</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:145px;">12-552-3</td>
			<td style="width:179px;">3.1.6)</td>
		</tr>
		<tr height="81">
			<td height="81" style="height:81px;width:303px;">Invertebrate iridescent virus 6 (IIV6)</td>
			<td style="width:288px;">Dr. Teixeria, L.</td>
			<td style="width:145px;"></td>
			<td>4) PLoS Biol, 6 (12), 2753-2763, doi: 10.1371/journal.pbio.1000002, (2008)</td>
		</tr>
		<tr height="141">
			<td height="141" style="height:141px;width:303px;">Listeria monocytogenes</td>
			<td style="width:288px;">ATCC</td>
			<td style="width:145px;">strain: 10403S</td>
			<td>4) Listeria monocytogenes strain 10403S (Bishop and Hinrichs, 1987) was grown in Difco Brain-heart infusion (BHI) broth (BD Biosciences) containing 50 &#181;g/ml streptomycin at 30 &#176;C.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:303px;">DNase I</td>
			<td style="width:288px;">Thermo Fisher Scientific(Invitrogen)</td>
			<td style="width:145px;">18068015</td>
			<td>gDNA degradation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">cDNA Synthesis Kit</td>
			<td style="width:288px;">Bio-Rad</td>
			<td style="width:145px;">1708891</td>
			<td>cDNA synthesis</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:303px;">IIV6_193R_F</td>
			<td style="width:288px;">IDT</td>
			<td style="width:145px;"></td>
			<td>qRT-PCR, 5&#39;- TCT TGT TTT CAG AAC CCC ATT -3&#39;</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:303px;">IIV6_193R_R</td>
			<td style="width:288px;">IDT</td>
			<td style="width:145px;"></td>
			<td>qRT-PCR, 5&#39;- CAC GAA GAA TGA CCA CAA GG -3&#39;</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:303px;">RpII_qRTPCR_fwd</td>
			<td style="width:288px;">SIGMA-ALDRICH</td>
			<td style="width:145px;"></td>
			<td>qRT-PCR, 5&#39;- GAA GCG TTT CTC CAA ACG -AG</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:303px;">RpII_qRTPCR_rev</td>
			<td style="width:288px;">SIGMA-ALDRICH</td>
			<td style="width:145px;"></td>
			<td>qRT-PCR, 5&#39;- TTG AGC GTA AGC ATC ACC -TG</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">SYBR Green qRT-PCR reagent</td>
			<td style="width:288px;">Thermo Fisher Scientific</td>
			<td style="width:145px;">K0251, K0252, K0253</td>
			<td>qRT-PCR</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:303px;">Real-Time PCR System</td>
			<td style="width:288px;">Thermo Fisher Scientific</td>
			<td style="width:145px;">4351107, 7500 Software v2.0</td>
			<td>qRT-PCR</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Anti-Listeria monocytogenes antibody</td>
			<td style="width:288px;">abcam</td>
			<td style="width:145px;">ab35132</td>
			<td>Western blot</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Anti-Actin antibody produced in rabbit</td>
			<td style="width:288px;">SIGMA-ALDRICH</td>
			<td style="width:145px;">A2066</td>
			<td>Western blot</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Anti-Rabbit IgG (H+L), HRP Conjugate</td>
			<td style="width:288px;">Promega</td>
			<td style="width:145px;">W4011</td>
			<td>Western blot</td>
		</tr>
	</tbody>
</table>

extraction of hemocytes from drosophila melanogaster larvae for microbial infection and analysis

Drosophila melanogaster, patojenik enfeksiyon sırasında hemocytes enfeksiyon boyunca bağışıklık yanıtındaki önemli bir rol oynamaktadır. Böylece, bu iletişim kuralının amacı belirli bir bağışıklık yerde uçar, yani hemocytes patojen işgali görselleştirmek için bir yöntem geliştirmektir. Burada 3 × 10 anlatılan yöntemle6 canlı hemocytes 200 Drosophila 3rd INSTAR larva ex vivo enfeksiyon için 30 dk içinde elde edilebilir. Alternatif olarak, hemocytes enfekte vivo içinde 3rd INSTAR larva için 24 saat sonrası enfeksiyon hemocyte çıkarma tarafından takip enjeksiyon yoluyla olabilir. Bu virüslü primer hücre tespit edildi, lekeli ve confocal mikroskobu kullanarak yansıma. Sonra 3D temsilcilikleri patojen işgal kesin olarak göstermek için görüntülerden üretildi. Ayrıca, yüksek kaliteli RNA qRT-PCR patojen mRNA şu tespiti için elde edilebilir enfeksiyon ve yeterli protein hulâsa Western blot analizi için bu hücrelerden. Birlikte ele alındığında, biz patojen işgalinin kesin uzlaşma ve enfeksiyon bakteriyel ve viral patojen türleri ve verimli bir yöntem hemocyte çıkarılması için yeterli canlı hemocytes Drosophila elde etmek için kullanarak onay için bir yöntem mevcut larva ex vivo ve in vivo enfeksiyon deneyler için.

Hemocytes ex vivo enfeksiyonu, 3 × 10 toplamak yaşamak için6 hemocytes 200 Drosophila 3rd INSTAR larvaları elde. Bizim yöntem geliştirmek için bir takım farklı teknikler denendi. Bireysel larva diseksiyon en fazla 1,5 saat sürer ve ortalama ~ 8000 hücre elde edilen en çok hangi koleksiyonun sonuna tarafından canlı değildi bu yöntemi18, kullanarak. Daha sonra hemocytes bulunan hemolymph çıkarmak için çalı?...

Watch this Scientific Journal Video about Extraction of Hemocytes from Drosophila melanogaster Larvae for Microbial Infection and Analysis at JoVE.com

Extraction of Hemocytes from Drosophila melanogaster Larvae for Microbial Infection and Analysis

Bu yöntem, patojen işgali üç boyutlu (3D) modelleri ile böcek hücreye görselleştirmek gösterilmiştir. Hemocytes Drosophila larvaları üzerinden viral veya bakteriyel patojenler, ex vivo veya vivo içindeile bulaşmış. Virüslü hemocytes daha sonra sabit ve görüntüleme için bir confocal mikroskop ve sonraki 3D hücresel yeniden yapılanma ile lekeli.

Hemocytes mikrobiyal enfeksiyon ve çözümleme çekme--dan Drosophila melanogaster larva

During the pathogenic infection of Drosophila melanogaster, hemocytes play an important role in the immune response throughout the infection. Thus, the ...

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JoVE Journal

Immunology and Infection

Extraction of Hemocytes from Drosophila melanogaster Larvae for Microbial Infection and Analysis

A Protocol for Collecting and Staining Hemocytes from the Yellow Fever Mosquito Aedes aegypti

A Protocol for Collecting and Staining Hemocytes from the Yellow Fever Mosquito Aedes aegypti

Yellow Fever Sivrisinek Hemocytes Toplama ve Boyama için Protokol Aedes aegypti</em

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A Simple Chelex Protocol for DNA Extraction from Anopheles spp.

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DNA ekstraksiyonu için A Simple Chelex Protokolü<em&gt; Anofel</em&gt; Spp.

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Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species

Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species

Toplam Protein Ekstraksiyon ve 2-D Jel Elektroforez Yöntemleri<em&gt; Burkholderia</em&gt; Türler

The investigation of  the intracellular protein levels of bacterial species is of importance to  understanding the pathogenic mechanisms of diseases ...

Systemic Bacterial Infection and Immune Defense Phenotypes in Drosophila Melanogaster

Systemic Bacterial Infection and Immune Defense Phenotypes in Drosophila Melanogaster

Sistemik Bakteriyel Enfeksiyon ve Bağışıklık Savunma FENOTİPİ içinde<em&gt; Drosophila melanogaster</em

The fruit fly Drosophila melanogaster is one of the premier model organisms for studying the function and evolution of immune defense. Many aspects of ...

Paraffin Embedding and Thin Sectioning of Microbial Colony Biofilms for Microscopic Analysis

Parafin gömme ve ince mikrobiyal koloni biyofilmler mikroskobik analizlerin yapıldığı için kesit

Sectioning via paraffin embedding is a broadly established technique in eukaryotic systems. Here we provide a method for the fixation, embedding, and ...

Combining Analysis of DNA in a Crude Virion Extraction with the Analysis of RNA from Infected Leaves to Discover New Virus Genomes

Ham Virion ayıklama DNA analizi yeni virüs genleri keşfetmek için virüslü yaprakları gelen RNA analizi ile birleştirerek

This metagenome approach is used to identify plant viruses with circular DNA genomes and their transcripts. Often plant DNA viruses that occur in low ...

Establishment of Viral Infection and Analysis of Host-Virus Interaction in Drosophila Melanogaster

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Viral enfeksiyon ve analiz Drosophila Melanogaster ana bilgisayar virüs etkileşimin kurulması

Virus spreading is a major cause of epidemic diseases. Thus, understanding the interaction between the virus and the host is very important to extend our ...

The Galleria mellonella Waxworm Infection Model for Disseminated Candidiasis

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Galleria mellonella Waxworm enfeksiyon modeli Dissemine kandidiyazis için

Candida species are common fungal commensals of humans colonizing the skin, mucosal surfaces, and gastrointestinal tract. Under certain conditions, ...

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Vaccines are a 20th century medical marvel. They have dramatically reduced the morbidity and mortality caused by infectious diseases and contributed to a ...