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"> Förbereda Vidhäftning Chambers (dagen före injektionen / dissekering) </p><p class="jove_step"> Vidhäftning analys Chambers </p><p class="jove_content"> Om du har en vanlig sammansatt mikroskop (optisk lokaliserar ovanför scenen) är det ncecessary att göra en låg höjd vidhäftning kammare enligt följande. Om du använder den inverterade-optik mikroskop, sedan &quot;NUNC Lab-Tek Chambered Coverglass&quot; bara kan användas, och hoppa över steg # 1 - 2. </p><ol><li> Fäst en &quot;tryck-till-tätning kisel isola...

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		<table border="1">
		<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">Denna video visar den teknik som används för beredning av arrangör och djur explants pol från Xenopus laevis embryon, inklusive användningen av ögonbrynet kniv - en specialiserad dissektion verktyg gjorda av ens ögonbrynet. Protokollet för montering en vidhäftning analys ges också, som sonder för förekomst av viktiga adhesionsmolekyler finns på ytan arrangören eller djurceller pol som är kritiska för korrekt utveckling.</p>

Dissekering av Organizer och Animal explants Pole från Xenopus laevis Embryon och församlingen i en cell Adhesion analys

Research

JoVE Journal

Biology

Dissection of 6.5 dpc Mouse Embryos

Dissekering av 6,5 DPC musembryon

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Plastic Embedding and Sectioning of Xenopus laevis Embryos

Plast inbäddning och Sektionering av Xenopus laevis embryon

Obtaining Eggs from Xenopus laevis Females

Skaffa Ägg från Xenopus laevis Kvinnor

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

Upprättande och fraktionering av Xenopus laevis Egg Utdrag

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

In Vitro Nuclear Assembly Använda Fraktionerade Xenopus Egg Utdrag

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

Mikroinjektion av Xenopus laevis oocyter

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

Tissue Fastställande Använda Transplant Animal Cap (ACT) analys i<em> Xenopus laevis</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

Live-cell imaging och kvantitativ analys av embryonala epitelceller i<em> Xenopus laevis</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

Ansikts Transplantationer i<em&gt; Xenopus laevis</em&gt; Embryon

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

Dissekering av<em&gt; Xenopus laevis</em&gt; Neural Crest för<em&gt; In vitro</em&gt; Explantation kultur eller<em&gt; In vivo</em&gt; Transplantation

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