Click the back button on your browser window to return to the Weisblat Lab website
<br>
		<p>
		<h3>Hoechst 33258 Counterstaining</h3>
		<ul>
			<li>the Hoechst precipitates with PO4, the embryos must be therefore treated in PO4 free buffer. </li>
			<li>Hoechst stock concentration is 1 mg/ml => must be diluted to the right concentration in any PO4 free buffer (Example: 0.1M Tris, pH 7.5). </li>
			<li>Incubate for +/- 30 min. </li>
			<li>Rinse in the buffer that has been used to prepare the mounting solution (see mounting). </li>
		<h2>Mounting </h2>
			<ul>
			<li>Mounting is usually made in 70% glycerol (70% glycerol + 30% buffer)</li>
			</ul>
			Before observation of any fluorescent dyed embryos, they can be treated with n-propyl gallate, it slows the bleaching and hence increases the delay of observing time. These embryos can also be mounted in 80% (and above) glycerol, the pH is more basic which slows the bleaching. 
		</p>