Name: live imaging of drosophila melanogaster embryonic hemocyte migrations
Uploaded: 2010-02-12T18:30:00
Description: Watch this Scientific Journal Video about Live Imaging Of Drosophila melanogaster Embryonic Hemocyte Migrations at JoVE.com

Bu protokol, Paul Martin ve Antonio Jacinto laboratuvarları ile işbirliği içinde ve çalışma ile geliştirilmiştir. Biz Bloomington Menkul Kıymetler Merkezi sinek satırları paylaşmak için devam etmek için mükemmel servis ve Drosophila topluluk için teşekkür ederiz . BS BBSRC bir proje hibe ile finanse edilmektedir. WW, Wellcome Trust Kariyer Geliştirme Bursu tarafından finanse edilmektedir.

Hazırlık <ol><li> Hemocyte özel bir Gal4 sürücüsü (örneğin srp-Gal4 2) ve genetik olarak kodlanmış UAS kontrol (örneğin UAS-GFP) altında bir floresan muhabiri içeren uygun Drosophila hatları alın. Srp-Gal4 UAS-GMA 3 veya CRQ-Gal4 UAS-GFP 4, 5, homozigot Sinekler görüntüleme amaçlı (nb GMA moesin aktin bağlayıcı etki alanı için erimiş GFP) yararlıdır; bir tartışma için aşağıya bakınız Gal4 sürücüleri v...

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	<li>Brand, A. H., Perrimon, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+gene+expression+as+a+means+of+altering+cell+fates+and+generating+dominant+phenotypes.">Targeted gene expression as a means of altering cell fates and generating dominant phenotypes.</a> Development. 118, 401-415 (1993).</li><li>Bruckner, 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+PDGF/VEGF+receptor+controls+blood+cell+survival+in+Drosophila.">The PDGF/VEGF receptor controls blood cell survival in Drosophila.</a> Dev Cell. 7, 73-84 (2004).</li><li>Dutta, D., Bloor, J. W., Ruiz-Gomez, M., VijayRaghavan, K., Kiehart, D. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Real-time+imaging+of+morphogenetic+movements+in+Drosophila+using+Gal4-UAS-driven+expression+of+GFP+fused+to+the+actin-binding+domain+of+moesin.">Real-time imaging of morphogenetic movements in Drosophila using Gal4-UAS-driven expression of GFP fused to the actin-binding domain of moesin.</a> Genesis. 34, 146-151 (2002).</li><li>Stramer, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Live+imaging+of+wound+inflammation+in+Drosophila+embryos+reveals+key+roles+for+small+GTPases+during+in+vivo+cell+migration.">Live imaging of wound inflammation in Drosophila embryos reveals key roles for small GTPases during in vivo cell migration.</a> J Cell Biol. 168, 567-573 (2005).</li><li>Wood, W., Jacinto, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila+melanogaster+embryonic+haemocytes:+masters+of+multitasking.">Drosophila melanogaster embryonic haemocytes: masters of multitasking.</a> Nat Rev Mol Cell Biol. 8, 542-551 (2007).</li><li>Halfon, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+fluorescent+protein+reporters+for+use+with+the+Drosophila+Gal4+expression+system+and+for+vital+detection+of+balancer+chromosomes.">New fluorescent protein reporters for use with the Drosophila Gal4 expression system and for vital detection of balancer chromosomes.</a> Genesis. 34, 135-138 (2002).</li><li>Sullivan, W., Ashburner, M., Hawley, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Drosophila protocols. , (2000).</li><li>Tepass, U., Fessler, L. 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Bu prosedürün en önemli unsurlardan açıkça etiketli hemocytes sağlıklı embriyoların seçimi ve onlara zarar vermeden bunları dikkatli bir şekilde monte etmek için. Birkaç saat süreyle imaged embriyoların Halokarbon yağı sonra dehidratasyon dayanıklı ve bir kez monte edilebilir. Bizim ellerde 40X amacı ile her üç dakikada bir Zeiss LSM510 konfokal mikroskop görüntüleri z yığını alarak, embriyo ya da açık foto-hasar ihmal dehidratasyon üç saat görüntü hemocytes,. Hemocytes son derece d...

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<td>Cell strainer</td>
<td>&nbsp;</td>
<td>BD Falcon</td>
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<td>70&mu;m pores</td>
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<td>Halcarbon oil 700</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>H8898</td>
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<td>Lumox/Petriperm dish</td>
<td>&nbsp;</td>
<td>Sarstedt</td>
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live imaging of drosophila melanogaster embryonic hemocyte migrations

Birçok çalışma, adres hücre göçü<em&gt; In vitro</em&gt; Yöntemleri, fizyolojik ilgili çevre ise organizmanın kendini. Burada montaj için bir protokol<em&gt; Drosophila melanogaster</em&gt; Embriyo ve floresan etiketli hemocytes sonraki canlı görüntüleme, bu organizmanın embriyonik makrofajlar. Gal4 UAS sistemini kullanarak<sup&gt; 1</sup&gt; Embriyo boyunca gelişimsel dağılma takip hemocytes genetik olarak kodlanmış, floresan etiketli işaretleyicileri çeşitli ifadesi sürücü. Embriyo gelişiminin istenilen aşamada aşağıdaki toplama, dış koryon kaldırılır ve embriyoların daha sonra bir hidrofobik, gaz geçirgen bir zar ve canlı görüntüleme için bir cam lamel arasında Halokarbon yağ monte edilmiştir. Hız ve yönlülük, F-aktin ve mikrotübüller bu sitoskeletal bileşenlerin dinamikleri ile ilgili daha detaylı bilgi verebilir floresan gazetecilere kullanımı ile birleştiğinde yüksek çözünürlüklü görüntüleme gibi brüt göçmen parametrelere ek olarak.

Watch this Scientific Journal Video about Live Imaging Of Drosophila melanogaster Embryonic Hemocyte Migrations at JoVE.com

Live Imaging Of Drosophila melanogaster Embryonic Hemocyte Migrations

Drosophila hemocytes gelişmekte olan embriyonun bütünlüğü içinde dağılırlar . Bu protokol, floresan etiketli hemocytes embriyoları kullanılarak bu göçler montaj ve görüntü nasıl gösteriyor.

Uygulamalarda Canlı Görüntüleme Drosophila melanogaster Embriyonik Hemocyte Göçler

Many studies address cell migration using in vitro methods, whereas the physiologically relevant environment is that of the organism itself. Here we ...

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Live Imaging Of Drosophila melanogaster Embryonic Hemocyte Migrations

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