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

このプロトコルは、ポールマーティンとアントニオサンジャシントの研究室とのコラボレーションの中でとの我々の仕事を通じて開発されています。我々は、フライラインを共有し続けるための素晴らしいサービスとショウジョウバエコミュニティのためのブルーミントンストックセンターに感謝します。 BSは現在BBSRCプロジェクトの助成金によって賄われています。 WWは、ウェルカムトラストのキャリア開発奨学金によって運営されている。

準備<ol><li>血球特異Gal4のドライバ（例：SRP - Gal4の 2）とUAS制御（例えば、UAS - GFP）の下で遺伝的に符号化された蛍光レポーターを含む適切なショウジョウバエの行を取得します。 SRP - GAL4、UAS - GMA 3またはCRQ - GAL4、UAS - GFP 4、5のホモ接合体ハエは、イメージングのために（NB GMAはモエシンのアクチン結合ドメインに融合したGFPです）...

<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. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Condensation+of+the+central+nervous+system+in+embryonic+Drosophila+is+inhibited+by+blocking+hemocyte+migration+or+neural+activity.">Condensation of the central nervous system in embryonic Drosophila is inhibited by blocking hemocyte migration or neural activity.</a> Dev Biol. 279, 233-243 (2005).</li><li>Paladi, M., Tepass, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Function+of+Rho+GTPases+in+embryonic+blood+cell+migration+in+Drosophila.">Function of Rho GTPases in embryonic blood cell migration in Drosophila.</a> J Cell Sci. 117, 6313-6326 (2004).</li><li>Vlisidou, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila+embryos+as+model+systems+for+monitoring+bacterial+infection+in+real+time.">Drosophila embryos as model systems for monitoring bacterial infection in real time.</a> PLoS Pathog. 5, e1000518-e1000518 (2009).</li><li>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=Dynamic+actin-based+epithelial+adhesion+and+cell+matching+during+Drosophila+dorsal+closure.">Dynamic actin-based epithelial adhesion and cell matching during Drosophila dorsal closure.</a> Curr Biol. 10, 1420-1426 (2000).</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=Imaging+cell+movement+during+dorsal+closure+in+Drosophila+embryos.">Imaging cell movement during dorsal closure in Drosophila embryos.</a> Methods Mol Biol. 294, 203-210 (2005).</li><li>Kunwar, P. 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この手順の最も重要な要素は明記された血球との健全な胚の選択であり、それらを傷つけることなく慎重にそれらをマウントする。一度胚は、脱水に耐性があると一度にマウントされている数時間のためにイメージングすることができるハロカーボンのオイルになります。私たちの手では40倍の目標と私たちのツァイスLSM510共焦点顕微鏡で3分ごとに画像のz -スタックを取っ胚やフォトダメー?...

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<td>Cell strainer</td>
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live imaging of drosophila melanogaster embryonic hemocyte migrations

使用して多くの研究のアドレスの細胞遊走<em&gt; in vitroで</em&gt;の方法は、生理学的に関連する環境に対し、生物自体のことです。ここでは、取り付けのためのプロトコルを提示<em&gt;キイロショウジョウバエ</em&gt;胚と蛍光標識血球のその後のライブイメージング、この生物の胚マクロファージ。 GAL4 - UASシステムを使用する<sup&gt; 1</sup&gt;我々は胚全体に彼らの発達分散に従うことを血球に遺伝的に符号化された、蛍光標識マーカーの様々な発現を駆動する。開発の目的の段階で胚のコレクションに続いて、外側の絨毛膜が除去され、胚は、その後疎水性、ガス透過膜とライブイメージング用ガラスカバースリップ間のハロカーボンのオイルにマウントされます。このような速度と方向性として総渡り鳥のパラメータに加えて、蛍光F -アクチンの記者と微小管の使用と相まって、高解像度のイメージングは​​、これらの細胞骨格成分の動態に関するより詳細な情報を提供することができます。

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

ショウジョウバエの血球は胚の全体にわたって分散させる。このプロトコルは、蛍光標識した血球で胚を使用してこれらの移行をマウントし、イメージする方法を示します。

のライブイメージングキイロショウジョウバエ胚性血球移行

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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Biology

Live Imaging Of Drosophila melanogaster Embryonic Hemocyte Migrations

Dissection of Larval CNS in Drosophila Melanogaster

キイロショウジョウバエの幼虫中枢神経系の解剖

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

Live Imaging of the Zebrafish Embryonic Brain by Confocal Microscopy

共焦点顕微鏡によるゼブラフィッシュ胚の脳のイメージングを生きる

In this video, we demonstrate the method our lab has developed to analyze the cell shape changes and rearrangements required to bend and fold the ...

Live-cell Imaging and Quantitative Analysis of Embryonic Epithelial Cells in Xenopus laevis

Live-cell Imaging and Quantitative Analysis of Embryonic Epithelial Cells in Xenopus laevis

生細胞イメージングとの胚上皮細胞の定量分析アフリカツメガエル</em

Embryonic epithelial cells serve as an ideal model to study morphogenesis where multi-cellular tissues undergo changes in their geometry, such as changes ...

Dissection of Oenocytes from Adult Drosophila melanogaster

Dissection of Oenocytes from Adult Drosophila melanogaster

大人からOenocytesの解剖キイロショウジョウバエ</em

In Drosophila melanogaster, as in other insects, a waxy layer on the outer surface of the cuticle, composed primarily of hydrocarbon compounds, provides ...

Isolation of Drosophila melanogaster Testes

Isolation of Drosophila melanogaster Testes

の分離キイロショウジョウバエ精巣

The testes of Drosophila melanogaster provide an important model for the study of stem cell maintenance and differentiation, meiosis, and soma-germline ...

Preparing Individual Drosophila Egg Chambers for Live Imaging

Preparing Individual Drosophila Egg Chambers for Live Imaging

個々の準備ショウジョウバエ</emライブイメージングのための>エッグチェンバース

Live cell imaging is an important technique applied to a number of Drosophila tissues used as models to investigate topics such as axis specification, ...

Live Imaging of GFP-labeled Proteins in Drosophila Oocytes

Live Imaging of GFP-labeled Proteins in Drosophila Oocytes

でGFP標識タンパク質のイメージングを生きる<em&gt;ショウジョウバエ</em&gt;卵母細胞

The  Drosophila oocyte has been established as a versatile system for investigating  fundamental questions such as cytoskeletal function, cell ...

Measurement of Lifespan in Drosophila melanogaster

Measurement of Lifespan in Drosophila melanogaster

における寿命の測定<em&gt;キイロショウジョウバエ</em

Aging is a phenomenon that results in steady physiological deterioration in nearly all organisms in which it has been examined, leading to reduced ...

Live Imaging of Apoptotic Cell Clearance during Drosophila Embryogenesis

Live Imaging of Apoptotic Cell Clearance during Drosophila Embryogenesis

アポトーシス細胞のクリアランスのイメージング中に生きる<em&gt;ショウジョウバエ</em&gt;胚

The  proper elimination of unwanted or aberrant cells through apoptosis and  subsequent phagocytosis (apoptotic cell clearance) is crucial for normal  ...

Imaging Through the Pupal Case of Drosophila melanogaster

Imaging Through the Pupal Case of Drosophila melanogaster

の蛹ケースを画像化<em&gt;キイロショウジョウバエ</em

The longstanding use of Drosophila as a model for cell and developmental biology has yielded an array of tools. Together, these techniques have enabled ...