Biomod/2013/OSU/protocol/custom tem grids

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<img src="http://openwetware.org/images/1/15/Ohiomod2013.jpg" height="60%" width="60%" style="margin-left:-40px;"> <div id="navbar" style="margin-left:auto; margin-right:auto;text-align:center;width:650px;height:auto;" > <ul id="homebutton" class="navigation-bar"> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU">Home</a></li> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/about">About</a> <ul> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/abstract">Abstract</a></li> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/goals">Goals</a></li>

<li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/about">Description</a> </ul> </li> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/experiments">Project</a> <ul> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/experiments">Life of Project</a></li> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/protocol">Protocols</a></li> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/labnotes">Lab Notes</a></li> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/meetingnotes">Meeting Notes</a></li> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/resources">Resources</a></li> </ul> </li> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/results">Results</a> <ul> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/results">Experimental Results</a></li> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/data">Data</a></li> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/futurework">Future Work</a></li> </li> </ul> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/media">Media</a> <ul> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/projectvideo">Project Video</a><li> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/animations">Animations</a><li> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/videoblogs">Video Blog</a></li> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/photos">Photos</a></li> </ul> </li> <li><a href="http://openwetware.org/wiki/Biomod/2013/OSU/team">Team</a></li> </ul>

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                       <a href="http://openwetware.org/wiki/Biomod/2013/OSU/protocol/folding_reaction">

<img src="http://openwetware.org/images/9/9e/Protocol_01.gif" width="129" height="125" alt=""></a></td> <td rowspan="2" align="left" valign="top"> <a href="http://openwetware.org/wiki/Biomod/2013/OSU/protocol/gel_prep"> <img src="http://openwetware.org/images/4/48/Protocol_02.gif" width="99" height="123" border="0" alt=""></a></td> <td align="left" valign="top"> <a href="http://openwetware.org/wiki/Biomod/2013/OSU/protocol/negative_staining"> <img src="http://openwetware.org/images/e/ed/Protocol_03.gif" width="116" height="122" border="0" alt=""></a></td> <td rowspan="2" align="left" valign="top"> <a href="http://openwetware.org/wiki/Biomod/2013/OSU/protocol/custom_tem_grids"> <img src="http://openwetware.org/images/a/ae/Protocol_04.gif" width="93" height="123" border="0" alt=""></a></td> <td colspan="2" rowspan="2" align="left" valign="top"> <a href="http://openwetware.org/wiki/Biomod/2013/OSU/protocol/drugloading"> <img src="http://openwetware.org/images/e/e9/Protocol_05.gif" width="132" height="123" border="0" alt=""></a></td> <td colspan="2" rowspan="2" align="left" valign="top"> <a href="http://openwetware.org/wiki/Biomod/2013/OSU/protocol/TIRF"> <img src="http://openwetware.org/images/f/fd/Protocol_06.gif" width="137" height="123" border="0" alt=""></a></td> <td rowspan="2" align="left" valign="top"> <a href="http://openwetware.org/wiki/Biomod/2013/OSU/protocol/MTT"> <img src="http://openwetware.org/images/2/20/Protocol_07.gif" width="94" height="123" border="0" alt=""></a></td> <td> <img src="http://openwetware.org/images/5/52/Spacer.gif" width="1" height="122" alt=""></td> </tr> <tr> <td rowspan="3"> <img src="http://openwetware.org/images/0/0b/Protocol_08.gif" width="116" height="3" alt=""></td> <td> <img src="http://openwetware.org/images/5/52/Spacer.gif" width="1" height="1" alt=""></td> </tr> <tr> <td rowspan="2"> <img src="http://openwetware.org/images/5/59/Protocol_09.gif" width="99" height="2" alt=""></td> <td colspan="2" rowspan="2"> <img src="http://openwetware.org/images/e/e4/Protocol_10.gif" width="211" height="2" alt=""></td> <td colspan="2" align="left" valign="top"> <a href="http://openwetware.org/wiki/Biomod/2013/OSU/protocol/cellanalysis"> <img src="http://openwetware.org/images/3/3c/Protocol_11.gif" width="108" height="1" border="0" alt=""></a></td> <td colspan="2" rowspan="2"> <img src="http://openwetware.org/images/8/83/Protocol_12.gif" width="137" height="2" alt=""></td> <td> <img src="http://openwetware.org/images/5/52/Spacer.gif" width="1" height="1" alt=""></td> </tr> <tr> <td colspan="2"> <img src="http://openwetware.org/images/7/77/Protocol_13.gif" width="108" height="1" alt=""></td> <td> <img src="http://openwetware.org/images/5/52/Spacer.gif" width="1" height="1" alt=""></td> </tr> <tr> <td> <img src="http://openwetware.org/images/5/52/Spacer.gif" width="129" height="1" alt=""></td> <td> <img src="http://openwetware.org/images/5/52/Spacer.gif" width="99" height="1" alt=""></td> <td> <img src="http://openwetware.org/images/5/52/Spacer.gif" width="116" height="1" alt=""></td> <td> <img src="http://openwetware.org/images/5/52/Spacer.gif" width="93" height="1" alt=""></td> <td> <img src="http://openwetware.org/images/5/52/Spacer.gif" width="118" height="1" alt=""></td> <td> <img src="http://openwetware.org/images/5/52/Spacer.gif" width="14" height="1" alt=""></td> <td> <img src="http://openwetware.org/images/5/52/Spacer.gif" width="94" height="1" alt=""></td> <td> <img src="http://openwetware.org/images/5/52/Spacer.gif" width="43" height="1" alt=""></td> <td> <img src="http://openwetware.org/images/5/52/Spacer.gif" width="94" height="1" alt=""></td> <td></td> </tr> </table> </center>

<div id="protocol-content" style="width:65%;margin-left:auto;margin-right:auto;"> <h1>Preparing Custom Grids for TEM</h1>

<h2>Coat grids with a thin collodion plastic film</h2> <h3>Materials</h3> <ul> <li>Collodion</li> <li>minimum 20 cm diameter bowl/basin</li> <li>TEM copper grids 200 or 400</li> <li>vellum paper (Pergament-Papier)</li> <li>tweezers, forceps</li> <li>razor blade</li> <li>200 pipet</li> </ul>

<h3>Procedure</h3> <ul> <li>Completely fill basin with ddH2O right to the rim.</li> <li>Use razor blade to shorten a 200er pipet tip to create a 3mm diameter aperture.</li> <li>Pipet out 50 μL collodion solution.</li> <li>Eject the collodion from approx 10 cm onto the calm water surface. CAUTION: collodion is toxic! </li> <li>Ensure proper ventilation and hold breath while handling the collodion!</li> <li>A collodion film will spread across the water surface and harden. Wait for 5 min.</li> <li>Use forcepts to position copper grids onto the collodion film. Put shiny (specular-reflecting) side of the grids facing the collodion. Arrange in a neat hexagonal or square pattern.</li> <li>Position vellum paper on top of girds. Let soak with water.</li> <li>Put collodion film around the circumference of the vellum paper by using the razor blade.</li> <li>Carefully pick up the vellum paper at one corner/edge and pull out of the water.</li> <li>Turn the vellum paper with the grids facing up, put into petri dish, partially cover and let dry overnight in a clean dust free area.</li> <li>Ready!</li> </ul>

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