<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. ...

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<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 Views.  University of California, Irvine (UCI). Dimostrato in questo video sono le tecniche per flash congelamento e sezionamento del tessuto embrionale del cervello di topo. Consigli utili per l'utilizzo del criostato sono dati, compresi i metodi di risoluzione dei problemi che possono essere utilizzati durante il taglio per assicurare che le sezioni risultanti tessuti sono privi di crepe e altre distorsioni.