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			<h3>Injection</h3>
			<h2>Materials</h2>
			<ul>
				<li>Micropipettes and micropipette puller to make the needles for injections. </li>
				<li>3 ml aliquots of lineage tracers (concentration 100 mg/ml). Once taken out from the freezer, the tracers can be stored for at least a month at 4oC </li>
				<li>Fast green FCF 4% in 0.2 KCl N </li>
				<li>Filters and syringe to filter the fast green </li>
				<li>Oscilloscope and injector</li>
			</ul>

			<h2>Lineage tracers and fast green/phenol red </h2>
			 In order to see the injection of the tracer within the injected cells, the fast green or phenol red dye is added to the tracer for visual monitoring at the ratio of 1(dye): 3(tracer). Before being added, the fast green must be filtered, or, if not filtered, centrifuge mix after adding.

			<h2>Needles </h2>
			Once the fast green is added to the tracer, the needles can be filled by dipping the flat end in the tracer. Then the needle can be stored in a petri dish on top of "plasticine" so as to avoid any movement which may break them. 
			Unused filled needles can be stored at 4oC for at least two weeks.

			<h2>Injections</h2>
			After having been shown the injection process, if you still feel uncomfortable with the injections apparatus, ask someone in the lab who is familiar with the injection process.  If you have any doubt as you are manipulating the injection apparatus, ASK someone for help before you cause irreversible damage!! 
	

		 
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