Name: dissection of organizer and animal pole explants from xenopus laevis embryos and assembly of a cell adhesion assay
Uploaded: 2007-04-29T00:00:00
Description: Watch this Scientific Journal Video about Dissection of Organizer and Animal Pole Explants from Xenopus laevis Embryos and Assembly of a Cell Adhesion Assay at JoVE.com

<p class="jove_title">準備接着チェンバース（注入/解剖前日） </p><p class="jove_step">接着アッセイチャンバー</p><p class="jove_content">あなたが従来の複合顕微鏡を（ステージ上の光学位置）がある場合、それは低身長の付着室は、次のようにするためncecessaryです。倒立光学顕微鏡を使用している場合は、&quot;NUNCのLab - Tek社チェンカバーガラスは、&quot;単に使用され、ステップ＃1へスキップする - 2。 </p><ol><li> 40 × 24ミリ...

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		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>1x MBS-H</td>
<td>Buffer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Modified Barth&#x2019;s medium: 1L recipe
5.14g		NaCl
75mg		KCl
0.2g		NaHCO3
0.2g		MgSO4&#x2022;7H2O
60mg		CaCl2&#x2022;2H2O
80mg		Ca(NO3)2&#x2022;4H2O
2.38g		HEPES, free-acid
Adjust the pH to 7.4 with NaOH.  Autoclave.
</td>
</tr>
<tr>
<td>CMF-MBS	</td>
<td>Buffer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>This is Ca2+/Mg2+-free version of MBS-H. 500ml recipe :
2.57g		NaCl
37.5mg		KCl
0.1g		NaHCO3
1.19g		HEPES, free-acid
Adjust the pH to 7.4 with NaOH.  Autoclave.
</td>
</tr>
<tr>
<td>1% agarose/1x Barth&#x2019;s plates</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>as many as the number of samples</td>
</tr>
<tr>
<td>1% agarose/CMF-MBS plates</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>as many as the number of samples</td>
</tr>
<tr>
<td>E-Cadherin/Fc chimera </td>
<td>Reagent</td>
<td>Sigma</td>
<td>E-2278</td>
<td>Dissolve 0.1mg of solid E-cadherin in 100ul of 1x MBS-H containing 40% glycerol to make 100x master stock (1.0mg/ml).  Store it in &#x2013;20C.  Make 1/100 dilution with 1x MBS-H to make 10ug/ml working solution just before you use.  You need 200ul per samples of this working solution.</td>
</tr>
<tr>
<td>Fibronectin, 0.1% solution </td>
<td>Reagent</td>
<td>Sigma</td>
<td>F-1141</td>
<td>This pre-made solution can be used as 50x stock.  Make 1/50 dilution with 1x MBS-H make 20ug/ml working solution just before you use.  You need 200ul per samples of this working solution.
I do NOT recommend using solid Fibronectin, because it is difficult to dissolve at 0.1% concentration to make the stock solution. Although it is more expensive, this pre-made 0.1% solution is better to obtain consistent results.
</td>
</tr>
<tr>
<td>4% BSA /MBS-H</td>
<td>Buffer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>BSA: molecular biology grade fraction V</td>
</tr>
<tr>
<td>Sigmacoted yellow tips </td>
<td>Tool</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>No.1 cover slips</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>40x24mm or 60x24mm</td>
</tr>
<tr>
<td>Press-to-seal silicon insulator</td>
<td>Tool</td>
<td>Glace Bio-Labs</td>
<td>JTR20-2.5</td>
<td>2.5mm depth x 20mm diameter </td>
</tr>
<tr>
<td>NUNC Lab-Tek Chambered Coverglass</td>
<td>Tool</td>
<td>Fisher</td>
<td>12-565-471</td>
<td>2wells/slides </td>
</tr>
<tr>
<td>Grid slide glass</td>
<td>Tool</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>or Finder graticule for cell counting</td>
</tr>		</tbody>
		</table>
		</div>
	

dissection of organizer and animal pole explants from xenopus laevis embryos and assembly of a cell adhesion assay

Watch this Scientific Journal Video about Dissection of Organizer and Animal Pole Explants from Xenopus laevis Embryos and Assembly of a Cell Adhesion Assay at JoVE.com

Dissection of Organizer and Animal Pole Explants from Xenopus laevis Embryos and Assembly of a Cell Adhesion Assay

<p class="jove_content">自分の眉製の特殊な解剖ツール - このビデオでは、眉毛のナイフの使用を含むアフリカツメガエルの胚から主催者と動物極の外植片、の調製に用いられる技法を示しています。接着アッセイを組み立てるためのプロトコルも与え、その適切な開発のための重要な面の主催者や動物極細胞上に存在する主要な接着分子の存在のためのプローブです。</p>

アフリカツメガエルの胚および細胞接着アッセイのアセンブリから主催し、動物極の外植片の解剖

Research

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Dissection of 6.5 dpc Mouse Embryos

6.5 DPCのマウス胚の解剖

Analysis of gene expression patterns during early stages of mammalian embryonic development can provide important clues about gene function, cell-cell ...

Plastic Embedding and Sectioning of Xenopus laevis Embryos

プラスチック製の埋め込みとアフリカツメガエル胚のセクショニング

Obtaining Eggs from Xenopus laevis Females

アフリカツメガエル女性から卵子を入手

The eggs of Xenopus laevis intact, lysed, and/or fractionated are useful for a wide variety of experiments. This protocol shows how to induce egg laying, ...

Preparation and Fractionation of Xenopus laevis Egg Extracts

アフリカツメガエル卵抽出物の調製と分画

Crude and fractionated Xenopus egg extracts can be used to provide ingredients for reconstituting cellular processes for morphological and biochemical ...

In Vitro Nuclear Assembly Using Fractionated Xenopus Egg Extracts

分別アフリカツメガエルの卵抽出を用いたin vitro核アセンブリで

Nuclear membrane assembly is an essential step in the cell division cycle; this process can be replicated in the test tube by combining Xenopus sperm ...

Microinjection of Xenopus Laevis Oocytes

アフリカツメガエル卵母細胞のマイクロインジェクション

Microinjection of Xenopus laevis oocytes followed by thin-sectioning electron microscopy (EM) is an excellent system for studying nucleocytoplasmic ...

Tissue Determination Using the Animal Cap Transplant (ACT) Assay in <em>Xenopus laevis</em>

Tissue Determination Using the Animal Cap Transplant ACT Assay in Xenopus laevis

でアニマルキャップの移植（ACT）アッセイを用いて組織の決定<em>アフリカツメガエル</em

Many proteins play a dual role in embryonic development. Those that regulate cell fate determination in a specific tissue can also affect the development ...

Live-cell Imaging and Quantitative Analysis of Embryonic Epithelial Cells in <em>Xenopus laevis</em>

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<em>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 ...

Facial Transplants in <em>Xenopus laevis</em> Embryos

Facial Transplants in Xenopus laevis Embryos

顔面移植<em&gt;アフリカツメガエル</em&gt;胚

Craniofacial birth defects occur in 1 out of every 700 live births, but etiology is rarely known due to limited understanding of craniofacial development. ...

Dissection of <em>Xenopus laevis</em> Neural Crest for <em>in vitro</em> Explant Culture or <em>in vivo</em> Transplantation

Dissection of Xenopus laevis Neural Crest for in vitro Explant Culture or in vivo Transplantation

の解剖<em&gt;アフリカツメガエル</em&gt;神経堤のため<em&gt; in vitroで</em&gt;外植片培養又は<em&gt; in vivoで</em&gt;移植

The neural crest (NC) is a transient dorsal neural tube cell population that undergoes an epithelium-to-mesenchyme transition (EMT) at the end of ...