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"> Vorbereitung Adhesion Chambers (der Tag vor der Injektion / Dissektion) </p><p class="jove_step"> Adhesion Assay Chambers </p><p class="jove_content"> Wenn Sie eine herkömmliche zusammengesetzte Mikroskop (optisch lokalisiert über der Bühne) haben, ist es ncecessary um eine niedrige Bauhöhe Haftung Kammer wie folgt. Bei Verwendung des invertierten-Optik Mikroskop, dann &quot;NUNC Lab-Tek Chambered Deckglas&quot; kann nur verwendet werden, und überspringen Sie die Schritte # 1 bis 2. </p><ol><li> Bringen Sie eine der ...

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		<thead>
		<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">Dieses Video zeigt die Technik für die Herstellung von Veranstalter und animalen Pol Explantaten aus Xenopus laevis Embryonen, einschließlich der Verwendung der Augenbraue Messer eingesetzt - ein spezialisiertes Werkzeug Dissektion einer Augenbraue gemacht. Das Protokoll zur Montage eines Adhäsionsassay ist auch gegeben, die Sonden für die Präsenz der wichtigsten Adhäsionsmoleküle auf der Oberfläche Veranstalter oder animalen Pol Zellen, die entscheidend für die richtige Entwicklung vorhanden sind.</p>

Präparation Organizer und Animal Pole Explantate aus Xenopus laevis Embryos und Montage eines Cell Adhesion Assay

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Microinjection of Xenopus Laevis Oocytes

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

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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

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

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

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