http://2007.igem.org/wiki/index.php?title=Paris/July_12&feed=atom&action=historyParis/July 12 - Revision history2024-03-28T15:06:05ZRevision history for this page on the wikiMediaWiki 1.16.5http://2007.igem.org/wiki/index.php?title=Paris/July_12&diff=23698&oldid=prevNicolas C. at 17:45, 7 October 20072007-10-07T17:45:28Z<p></p>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">[[Paris/July 11|yesterday]] -- [[Paris/July 13|tomorrow]] <br></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Preparation of a stock of phage on w121:==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Preparation of a stock of phage on w121:==</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>See [[Paris/PROTOCOLS#Preparation_of_the_P1_stock_on_the_w121_strain.|Protocols]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>See [[Paris/PROTOCOLS#Preparation_of_the_P1_stock_on_the_w121_strain.|Protocols]]</div></td></tr>
</table>Nicolas C.http://2007.igem.org/wiki/index.php?title=Paris/July_12&diff=17385&oldid=prevDavidpz: /* E.coli pKS::DGAT on LB oleate medium */2007-08-31T09:57:21Z<p><span class="autocomment">E.coli pKS::DGAT on LB oleate medium</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>We incubated E.coli pKS::DGAT overnight on different LB medium (see [[Paris/July_11|July 11]]).</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>We incubated E.coli pKS::DGAT overnight on different LB medium (see [[Paris/July_11|July 11]]).</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>We did not observe any colony. Probably because spread plates with liquid culture in the freezer for one week was not a good idea...</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>We did not observe any colony. Probably because spread plates with liquid culture <ins class="diffchange diffchange-inline">kept </ins>in the freezer for one week was not a good idea...</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>On minimum medium, it did not grow too because what we thought to be M9 medium was only agar...</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>On minimum medium, it did not grow too because what we thought to be M9 medium was only agar...</div></td></tr>
</table>Davidpzhttp://2007.igem.org/wiki/index.php?title=Paris/July_12&diff=17384&oldid=prevDavidpz: /* Preparation of a stock of phage on w121: */2007-08-31T09:55:34Z<p><span class="autocomment">Preparation of a stock of phage on w121:</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Preparation of a stock of phage on w121:==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Preparation of a stock of phage on w121:==</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">David B: </del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">See [[Paris/PROTOCOLS#Preparation_of_the_P1_stock_on_the_w121_strain</ins>.<ins class="diffchange diffchange-inline">|Protocols]]</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">* Filtration of the Cacl2 and MgSO4 stocks</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">* Launch a culture:</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">** 15ml LB</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">** 225µl Cacl2</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">** 450µl MgSO4</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">** 90µl DAP</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">** 150µl W121 from an ON culture</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">* Let grow until OD=0</del>.<del class="diffchange diffchange-inline">2 (~1h)</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">* Add 100µl P1 (stock 10<sup>10</sup>/ml)</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">* Incubate 2h</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">* Add 200µl Chloroform</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">* Vortex, centrifuge (15min @ 3000rpm)</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">* Recover the supernatant + add 200µl Chloroform = Stock keep at 4°C</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Titration of bacteriophages P1 ==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Titration of bacteriophages P1 ==</div></td></tr>
</table>Davidpzhttp://2007.igem.org/wiki/index.php?title=Paris/July_12&diff=11550&oldid=prevDavidpz: /* E.coli pKS::DGAT on LB oletate medium */2007-07-26T10:33:16Z<p><span class="autocomment">E.coli pKS::DGAT on LB oletate medium</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:Paris_Acineto_Colonies.jpg|500px|center]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:Paris_Acineto_Colonies.jpg|500px|center]]</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>== E.coli pKS::DGAT on LB <del class="diffchange diffchange-inline">oletate </del>medium ==</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>== E.coli pKS::DGAT on LB <ins class="diffchange diffchange-inline">oleate </ins>medium ==</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>We incubated E.coli pKS::DGAT on different LB medium (see [[Paris/July_11|July 11]]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>We incubated E.coli pKS::DGAT <ins class="diffchange diffchange-inline">overnight </ins>on different LB medium (see [[Paris/July_11|July 11]]<ins class="diffchange diffchange-inline">).</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">We did not observe any colony. Probably because spread plates with liquid culture in the freezer for one week was not a good idea...</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">On minimum medium, it did not grow too because what we thought to be M9 medium was only agar...</ins></div></td></tr>
</table>Davidpzhttp://2007.igem.org/wiki/index.php?title=Paris/July_12&diff=11549&oldid=prevDavidpz: /* E.coli pKS::DGAT on LB oletate medium */2007-07-26T10:28:12Z<p><span class="autocomment">E.coli pKS::DGAT on LB oletate medium</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>== E.coli pKS::DGAT on LB oletate medium ==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>== E.coli pKS::DGAT on LB oletate medium ==</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>We incubated E.coli pKS::DGAT on different LB medium (see [[Paris/<del class="diffchange diffchange-inline">July11</del>]]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>We incubated E.coli pKS::DGAT on different LB medium (see [[Paris/<ins class="diffchange diffchange-inline">July_11|July 11</ins>]]</div></td></tr>
</table>Davidpzhttp://2007.igem.org/wiki/index.php?title=Paris/July_12&diff=11548&oldid=prevDavidpz: /* Acinetobacter microscopy */2007-07-26T10:27:46Z<p><span class="autocomment">Acinetobacter microscopy</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:Paris_Acineto_Colonies.jpg|500px|center]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:Paris_Acineto_Colonies.jpg|500px|center]]</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">== E.coli pKS::DGAT on LB oletate medium ==</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">We incubated E.coli pKS::DGAT on different LB medium (see [[Paris/July11]]</ins></div></td></tr>
</table>Davidpzhttp://2007.igem.org/wiki/index.php?title=Paris/July_12&diff=11409&oldid=prevDavidpz: /* Acinetobacter microscopy */2007-07-25T14:47:15Z<p><span class="autocomment">Acinetobacter microscopy</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>We looked under 5X fluorescence microscopy the fluorescence of colonies of Acinetobacter incubated on LNMM Nile Red for 4-5 days:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>We looked under 5X fluorescence microscopy the fluorescence of colonies of Acinetobacter incubated on LNMM Nile Red for 4-5 days:</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Visualisation of colonies stained by Nile Red. It seems that only cells in the middle of colonies (i.e. "old" cells) are producing triglycerides whereas cells on the edges doesn't. <br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Visualisation of colonies stained by Nile Red. It seems that only cells in the middle of colonies (i.e. "old" cells) are producing triglycerides whereas cells on the edges doesn't. <ins class="diffchange diffchange-inline">Without Nile red, the colonies show no fluorescence.</ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:Paris_Acineto_Colonies.jpg|500px|center]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:Paris_Acineto_Colonies.jpg|500px|center]]</div></td></tr>
</table>Davidpzhttp://2007.igem.org/wiki/index.php?title=Paris/July_12&diff=11408&oldid=prevDavidpz: /* Acinetobacter microscopy */2007-07-25T14:46:34Z<p><span class="autocomment">Acinetobacter microscopy</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>We looked under 5X fluorescence microscopy the fluorescence of colonies of Acinetobacter incubated on LNMM Nile Red for 4-5 days:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>We looked under 5X fluorescence microscopy the fluorescence of colonies of Acinetobacter incubated on LNMM Nile Red for 4-5 days:</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Visualisation of colonies stained by Nile Red. It seems that only cells in the middle of colonies (i.e. "old" cells) are producing triglycerides whereas cells on the edges doesn't. <br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Visualisation of colonies stained by Nile Red. It seems that only cells in the middle of colonies (i.e. "old" cells) are producing triglycerides whereas cells on the edges doesn't. <br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>[[Image:Paris_Acineto_Colonies.jpg|500px|center<del class="diffchange diffchange-inline">]</del>]]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>[[Image:Paris_Acineto_Colonies.jpg|500px|center]]</div></td></tr>
</table>Davidpzhttp://2007.igem.org/wiki/index.php?title=Paris/July_12&diff=11407&oldid=prevDavidpz at 14:46, 25 July 20072007-07-25T14:46:22Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>We looked under 5X fluorescence microscopy the fluorescence of colonies of Acinetobacter incubated on LNMM Nile Red for 4-5 days:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>We looked under 5X fluorescence microscopy the fluorescence of colonies of Acinetobacter incubated on LNMM Nile Red for 4-5 days:</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">* left : Acinetobacter grown in Low Nitrogen Mineral medium (see Protocol)</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">Visualisation of colonies stained by </ins>Nile Red. <ins class="diffchange diffchange-inline">It seems that only cells in </ins>the <ins class="diffchange diffchange-inline">middle of colonies (i</ins>.<ins class="diffchange diffchange-inline">e. "old" </ins>cells<ins class="diffchange diffchange-inline">) </ins>are <ins class="diffchange diffchange-inline">producing </ins>triglycerides whereas cells on the <ins class="diffchange diffchange-inline">edges doesn't</ins>. <ins class="diffchange diffchange-inline"><br></ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">* right : Acinetobacter grow in Low Nitrogen Mineral medium with </del>Nile Red <del class="diffchange diffchange-inline">(see Protocol)</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>[[Image:<ins class="diffchange diffchange-inline">Paris_Acineto_Colonies</ins>.jpg|<ins class="diffchange diffchange-inline">500px|center]</ins>]]</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">Exposure time and scaling is </del>the <del class="diffchange diffchange-inline">same for both images</del>. <del class="diffchange diffchange-inline">Clearly, some </del>cells <del class="diffchange diffchange-inline">on the right </del>are <del class="diffchange diffchange-inline">stained by Nile Red, which is specific to </del>triglycerides<del class="diffchange diffchange-inline">, </del>whereas <del class="diffchange diffchange-inline">almost all </del>cells on the <del class="diffchange diffchange-inline">left are not stained. Tipically, the size of one cell is about 5µm</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>[[Image:<del class="diffchange diffchange-inline">Paris_Acineto_NileRed</del>.jpg|<del class="diffchange diffchange-inline">850px</del>]]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
</table>Davidpzhttp://2007.igem.org/wiki/index.php?title=Paris/July_12&diff=11406&oldid=prevDavidpz at 14:45, 25 July 20072007-07-25T14:45:21Z<p></p>
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<td colspan='2' style="background-color: white; color:black;">← Older revision</td>
<td colspan='2' style="background-color: white; color:black;">Revision as of 14:45, 25 July 2007</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>We looked under 5X fluorescence microscopy the fluorescence of colonies of Acinetobacter incubated on LNMM Nile Red for 4-5 days:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>We looked under 5X fluorescence microscopy the fluorescence of colonies of Acinetobacter incubated on LNMM Nile Red for 4-5 days:</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">* left : Acinetobacter grown in Low Nitrogen Mineral medium (see Protocol)</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">* right : Acinetobacter grow in Low Nitrogen Mineral medium with Nile Red (see Protocol).</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">Exposure time and scaling is the same for both images. Clearly, some cells on the right are stained by Nile Red, which is specific to triglycerides, whereas almost all cells on the left are not stained. Tipically, the size of one cell is about 5µm.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">[[Image:Paris_Acineto_NileRed.jpg|850px]]</ins></div></td></tr>
</table>Davidpz