Biomod/2014/Hokudai/MATERIAL&METHOD: Difference between revisions
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<p class="example1">Process</p> | |||
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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> | |||
② GTP-buffer and tubulin solution were mixed at the ratio of 1 : 4 (Final tubulin concentration: 112.5 µM).<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> | |||
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> | |||
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> | |||
⑥ Microtubules were observed by using fluorescence* microscope and polarizing microscope.<br> | |||
*That time we used TAMRA labeled tubulin. | |||
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<p><a class="image" title="team hokudai"><img alt="team hokudai" src="https://upload.wikimedia.org/wikipedia/commons/f/f5/%E7%AD%8B%E8%82%89%E8%8B%B1%E8%AA%9E%E7%89%88.png" width="450" height="450" border="0" /></a> | |||
<a class="image" title="team hokudai"><img alt="team hokudai" src="https://upload.wikimedia.org/wikipedia/commons/8/86/%E7%B8%AE%E3%82%80%E3%82%B5%E3%83%AB%E3%82%B3%E3%83%A1%E3%82%A2.gif" width="400" height="300" border="0" /></a></p> | |||
<p>(Figure1.Left:Structure of muscle,Right:contraction of sarcomeres)</p> | |||
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Revision as of 01:33, 29 August 2014
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<body> <p class="example1">Process</p> <p class="example6"> 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> ② GTP-buffer and tubulin solution were mixed at the ratio of 1 : 4 (Final tubulin concentration: 112.5 µM).<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> 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> 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> ⑥ Microtubules were observed by using fluorescence* microscope and polarizing microscope.<br>
- That time we used TAMRA labeled tubulin.
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<p><a class="image" title="team hokudai"><img alt="team hokudai" src="https://upload.wikimedia.org/wikipedia/commons/f/f5/%E7%AD%8B%E8%82%89%E8%8B%B1%E8%AA%9E%E7%89%88.png" width="450" height="450" border="0" /></a> <a class="image" title="team hokudai"><img alt="team hokudai" src="https://upload.wikimedia.org/wikipedia/commons/8/86/%E7%B8%AE%E3%82%80%E3%82%B5%E3%83%AB%E3%82%B3%E3%83%A1%E3%82%A2.gif" width="400" height="300" border="0" /></a></p> <p>(Figure1.Left:Structure of muscle,Right:contraction of sarcomeres)</p>
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