<ol>
<li>Fix tissue in 4% paraformaldehyde in PBS for desired time.</li>
<li>Sucrose infuse tissue (cryoprotection) <ol>
<li>Make 30% sucrose solution in PBS w/v in 2059 tube.</li>
<li>Rinse tissue 3x in PBS (~ 5 min with rocking).</li>
<li>Place tissue in 30% sucrose solution.&nbsp; Tissue will not sink.</li>
<li>Place the tissue in 4&deg;C overnight, or until it has sunk.</li>
</ol></li>
<li>Label appropriate size cryomold with information and orientation.</li>
<li>Fill cryomold with O.C.T. (avoid bubbles).</li>
<li>Transfer tissue to O.C.T. ...

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		<div>
		<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>Tissue-Tek Cryomold</td>
<td>&nbsp;</td>
<td>Ted Pella, Inc.</td>
<td>27181</td>
<td>&nbsp;</td>
<tr>
<td>O.C.T.</td>
<td>&nbsp;</td>
<td>Ted Pella, Inc</td>
<td>27050</td>
<td>&nbsp;</td>
<tr>
<td>Sucrose solution</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>30% sucrose solution in PBS w/v</td>
</tr>
<tr>
<td>paraformaldehyde</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>4% paraformaldehyde in PBS </td>
</tr>		</tbody>
		</table>
		</div>
			<div>
					</div>
		

flash freezing and cryosectioning e12.5 mouse brain

Watch this Scientific Journal Video about Flash Freezing and Cryosectioning E12.5 Mouse Brain at JoVE.com

Flash Freezing and Cryosectioning E12.5 Mouse Brain

<p class="jove_content">Demonstrated in this video are the techniques for flash freezing and sectioning embryonic brain tissue from mouse.  Useful tips for using the cryostat are given, including troubleshooting methods that can be used while cutting to ensure that the resultant tissues sections are free of cracks and other distortions.</p>

flash-freezing-and-cryosectioning-e125-mouse-brain

Research

JoVE Journal

Biology

42.3K visualizações.  University of California, Irvine (UCI). Demonstrado neste vídeo são as técnicas de congelamento instantâneo e secção tecido cerebral embrionário de rato. Dicas úteis para usar o criostato são dadas, incluindo métodos de solução de problemas que podem ser usados ​​durante o corte para garantir que as seções resultantes tecidos estão livres de rachaduras e outras distorções.