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

이 프로토콜은 폴 마틴과 안토니오 하신토의 연구소와 시간 및 공동 작업을 통해 우리의 개발되었습니다. 우리는 플라이 라인을 공유하는 지속에 대한 훌륭한 서비스와 Drosophila 커뮤니티에 대한 블루밍턴 주식 센터 주셔서 감사합니다. BS는 현재 BBSRC 프로젝트 부여하여 자금이다. WW는 웰컴 트러스트 경력 개발 원정대에 의해 후원됩니다.

준비 <ol><li> hemocyte 특정 Gal4 드라이버 (예 : SRP - Gal4 2)와 UAS 통제 (예 : UAS - GFP)에서 유전자 인코딩 형광 기자를 포함하는 적절한 Drosophila 라인을 구합니다. SRP - Gal4, UAS - GMA 3 crq - Gal4, UAS - GFP 4, 5 homozygous 파리 이미징을 위해 (NB GMA가 moesin의 굴지 바인딩 도메인에 융합 GFP)을 특히 유용합니다;의 토론 아래 참조 Gal4 드라이버와 UAS의 범위가 ...

<ol>
	<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. I., Aziz, A., Hartenstein, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Embryonic+origin+of+hemocytes+and+their+relationship+to+cell+death+in+Drosophila.">Embryonic origin of hemocytes and their relationship to cell death in Drosophila.</a> Development. 120, 1829-1837 (1994).</li><li>Millard, T. H., Martin, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+analysis+of+filopodial+interactions+during+the+zippering+phase+of+Drosophila+dorsal+closure.">Dynamic analysis of filopodial interactions during the zippering phase of Drosophila dorsal closure.</a> Development. 135, 621-626 (2008).</li><li>Doerflinger, H., Benton, R., Shulman, J. M., St Johnston, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+PAR-1+in+regulating+the+polarised+microtubule+cytoskeleton+in+the+Drosophila+follicular+epithelium.">The role of PAR-1 in regulating the polarised microtubule cytoskeleton in the Drosophila follicular epithelium.</a> Development. 130, 3965-3975 (2003).</li><li>Olofsson, B., Page, D. 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이 절차의 가장 중요한 요소는 명확하게 표시된 hemocytes과 건강한 태아의 선택이며, 그들을 손상하지 않고 신중하게 그들을 마운트합니다. 배아는 할로 카본 오일에 일단 그들이 탈수에 강한 있으며, 한번 장착은 몇 시간 동안 몇 군데하실 수 있습니다. 우리의 손에 우리는 40X 목적으로 매 삼분의 자이스 혈구 LSM510 공촛점 현미경에 이미지의 Z - 스택을 복용 배아 또는 사진 손상 분명 무시할 수 탈?...

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<td>Cell strainer</td>
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<td>BD Falcon</td>
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<td>Halcarbon oil 700</td>
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<td>Sigma</td>
<td>H8898</td>
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<td>Lumox/Petriperm dish</td>
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<td>Sarstedt</td>
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live imaging of drosophila melanogaster embryonic hemocyte migrations

사용하는 많은 연구 주소 셀 마이 그 레이션<em&gt; 체외에서</em&gt; 방법은 생리학 관련 환경 반면 유기체 자체이다. 여기서 우리는 장착의를위한 프로토콜을 제시<em&gt; Drosophila melanogaster</em&gt; 배아 및 찬란 분류 hemocytes의 후속 라이브 영상이 유기체의 배아 macrophages. Gal4 - UAS 시스템을 사용<sup&gt; 1</sup&gt; 우리는 배아를 통해 자신의 발달 분산에 따라 hemocytes에서 유전자 인코딩, 찬란 태그 마커의 다양한 표현을 운전. 개발의 원하는 단계에서 배아의 다음 컬렉션, 바깥쪽 chorion이 제거되고 배아는 다음 소수성, 가스 투과 막과 라이브 영상을위한 유리 coverslip 사이 할로 카본 기름에 마운트됩니다. 같은 속도와 방향, F - 굴지와 microtubules이 cytoskeletal 구성 요소의 역학에 관한 자세한 정보를 제공할 수의 형광 기자의 사용과 함께 높은 해상도의 영상으로 총 철새 매개 변수 이외에.

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

Drosophila의 hemocytes은 개발 배아의 전체를 통해 분산. 이 프로토콜은 찬란 표시된 hemocytes와 배아를 사용하여 이러한 마이 그 레이션을 마운트 및 이미지하는 방법을 보여줍니다.

라이브 영상 Drosophila melanogaster 배아 Hemocyte 마이 그 레이션

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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The central nervous system (CNS) of Drosophila larvae is complex and poorly understood.  One way to investigate the CNS is to use immunohistochemistry to ...

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