Biomod/2014/Hokudai/MATERIAL&METHOD: Difference between revisions

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<p><a1  class="image" title="team hokudai"> <a href="http://openwetware.org/wiki/Biomod/2014/Hokudai"><img alt="team hokudai" src="https://upload.wikimedia.org/wikipedia/commons/c/c0/Title%EF%BC%88%E4%BB%AE%EF%BC%89.png"  width="160" height="80" border="0" /></a></p>
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<nobr>
<p class="example111 example2 example3 example4 example5 clearLeft">  <a href="PROJECT">PROJECT</a>  <a href="DESIGN">DESIGN</a> <a>EXPERIMENTS&RESULTS</a> <a href="TEAM">TEAM</a> <a href="SPONSORS">SPONSORS</a>
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<p class="example7">Process</p>
<p class="example6"><br>① We prepared flow cells by placing a cover glass (18 × 18 mm2; MATSUNAMI) on an another slide glass (26 × 76 mm2) with a pair of double sided tapes as spacers.  Size of flow cell: 1.5 × 18 × 0.1 mm3 (~3 µL) (W × L × H) in dimension.</br>
<br>② GTP-buffer and tubulin solution were mixed at the ratio of 1 : 4 (Final tubulin concentration: 112.5 µM).</br>
<br>③ Tubulin mixture was incubated at 4 °C for 10 minutes to depolymerize it completely. It was confirmed that polymerization of tubulin had not been started yet by using a polarizing microscopes.</br>
<br>④ The backside of the area of interest in the flow cell was fixed with a black tape where polymerization of tubulin would be initiated by IR irradiation. </br>
<br>⑤ A part of the flow cell where black tape was fixed was warmed for 15 minutes by using IR irradiation (IRS-001C; IR SYSTEM Co., Ltd., 5V / 1.6A).</br>
<br>⑥ Microtubules were observed by using fluorescence* microscope and polarizing microscope.</br>
<br>*That time we used TAMRA labeled tubulin.</br>
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<p><a  class="image" title="team hokudai"><img alt="team hokudai" src="https://upload.wikimedia.org/wikipedia/commons/e/e1/IR%E6%9C%80%E7%B5%82%E7%89%88.JPG"  width="600" height="400" border="0" /></a>
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<p class="clearLeft">(Figure1:Iradiation to the flow cell by IR light)<br><br>
<p class="example7 clearLeft">Results</p>
<p><a  class="image" title="team hokudai"><img alt="team hokudai" src="https://upload.wikimedia.org/wikipedia/commons/e/e3/%E5%81%8F%E5%85%89%E9%A1%95%E5%BE%AE%E9%8F%A1.JPG"  width="450" height="400" border="0" /></a>
<a  class="image" title="team hokudai"><img alt="team hokudai" src="https://upload.wikimedia.org/wikipedia/commons/4/40/%E8%9B%8D%E5%85%89%E9%A1%95%E5%BE%AE%E9%8F%A1.jpg"  width="450" height="400" border="0" /></a></p>
<p class="clearLeft">(Figure2:Polarising microscopic(left),fluorescence microscopic
(right) image of well-oriented microtubules)<br>
Well-oriented microtubules was successfully formed under temparature
gradient using photo irradiation.
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Revision as of 10:34, 29 August 2014

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<p><a1 class="image" title="team hokudai"> <a href="http://openwetware.org/wiki/Biomod/2014/Hokudai"><img alt="team hokudai" src="https://upload.wikimedia.org/wikipedia/commons/c/c0/Title%EF%BC%88%E4%BB%AE%EF%BC%89.png" width="160" height="80" border="0" /></a></p>


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<nobr> <p class="example111 example2 example3 example4 example5 clearLeft"> <a href="PROJECT">PROJECT</a> <a href="DESIGN">DESIGN</a> <a>EXPERIMENTS&RESULTS</a> <a href="TEAM">TEAM</a> <a href="SPONSORS">SPONSORS</a> </nobr> </p>


<p class="example7">Process</p> <p class="example6"><br>① We prepared flow cells by placing a cover glass (18 × 18 mm2; MATSUNAMI) on an another slide glass (26 × 76 mm2) with a pair of double sided tapes as spacers. Size of flow cell: 1.5 × 18 × 0.1 mm3 (~3 µL) (W × L × H) in dimension.</br> <br>② GTP-buffer and tubulin solution were mixed at the ratio of 1 : 4 (Final tubulin concentration: 112.5 µM).</br> <br>③ Tubulin mixture was incubated at 4 °C for 10 minutes to depolymerize it completely. It was confirmed that polymerization of tubulin had not been started yet by using a polarizing microscopes.</br> <br>④ The backside of the area of interest in the flow cell was fixed with a black tape where polymerization of tubulin would be initiated by IR irradiation. </br> <br>⑤ A part of the flow cell where black tape was fixed was warmed for 15 minutes by using IR irradiation (IRS-001C; IR SYSTEM Co., Ltd., 5V / 1.6A).</br> <br>⑥ Microtubules were observed by using fluorescence* microscope and polarizing microscope.</br> <br>*That time we used TAMRA labeled tubulin.</br>

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<p><a class="image" title="team hokudai"><img alt="team hokudai" src="https://upload.wikimedia.org/wikipedia/commons/e/e1/IR%E6%9C%80%E7%B5%82%E7%89%88.JPG" width="600" height="400" border="0" /></a> </div> <p class="clearLeft">(Figure1:Iradiation to the flow cell by IR light)<br><br>

<p class="example7 clearLeft">Results</p>

<p><a class="image" title="team hokudai"><img alt="team hokudai" src="https://upload.wikimedia.org/wikipedia/commons/e/e3/%E5%81%8F%E5%85%89%E9%A1%95%E5%BE%AE%E9%8F%A1.JPG" width="450" height="400" border="0" /></a> <a class="image" title="team hokudai"><img alt="team hokudai" src="https://upload.wikimedia.org/wikipedia/commons/4/40/%E8%9B%8D%E5%85%89%E9%A1%95%E5%BE%AE%E9%8F%A1.jpg" width="450" height="400" border="0" /></a></p>

<p class="clearLeft">(Figure2:Polarising microscopic(left),fluorescence microscopic (right) image of well-oriented microtubules)<br> Well-oriented microtubules was successfully formed under temparature gradient using photo irradiation. </p>


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