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<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D">Hi Michael,<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D">Thanks for your reply and confirmation of the problem with hypoxia. We evaluate larvae from day 7 to 10 since we like to correlate imaging analysis with behavioral
response. Please keep me updated if you do solve the issue.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D">Best,<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D">Fredrik<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri","sans-serif";color:#1F497D"><o:p> </o:p></span></p>
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<p class="MsoNormal"><b><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif"">Från:</span></b><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif""> openspim-bounces@openspim.org [mailto:openspim-bounces@openspim.org]
<b>För </b>Michael Weber<br>
<b>Skickat:</b> den 27 jan</span><span lang="SV" style="font-size:10.0pt;font-family:"Tahoma","sans-serif"">uari 2016 15:44<br>
<b>Till:</b> openspim@openspim.org<br>
<b>Ämne:</b> Re: [OpenSPIM] survival FEP Tubes<o:p></o:p></span></p>
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<p class="MsoNormal">Hey Fredrik,<o:p></o:p></p>
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<p class="MsoNormal">thanks for watching our JoVE publication :). How old are the larvae you're trying to image? In our hands, FEP works great up to around 3 dpf. With older larvae we also see oxygen deficiency, because the tubes are gas-proof. Currently, those
developmental stages are not so interesting for us, so we did not spend much time on optimizing it. We have ideas though, e.g. putting holes in the tube to allow for oxygen diffusion.<o:p></o:p></p>
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<p class="MsoNormal"><o:p> </o:p></p>
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<p class="MsoNormal">Best,<o:p></o:p></p>
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<p class="MsoNormal">Michael<o:p></o:p></p>
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<p class="MsoNormal"><o:p> </o:p></p>
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<p class="MsoNormal">On Jan 27, 2016, at 2:51 PM, Fredrik Ek <<a href="mailto:fredrik.ek@med.lu.se">fredrik.ek@med.lu.se</a>> wrote:<o:p></o:p></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif"">Hi all,<o:p></o:p></span></p>
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<p style="margin:0cm;margin-bottom:.0001pt"><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif""> <o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif"">we have experienced some problems with survival of the larvae when mounted in the FEP tubes. They seems to suffocate after 1-2 h in the tube. Using the same agarose solution but
keeping the larvae covered on a glass slide they stay alive and healthy. We are following the protocol for long time imaging from JOVE (Weber et al 2014) but has also tested 0.5% and 1% low melting agarose solutions for embedding. It would be much appreciated
to get some input how to solve this issue.<o:p></o:p></span></p>
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<p style="margin:0cm;margin-bottom:.0001pt"><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif""> <o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif"">Thanks!<o:p></o:p></span></p>
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<p style="margin:0cm;margin-bottom:.0001pt"><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif""> <o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif"">Fredrik<o:p></o:p></span></p>
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<p style="margin:0cm;margin-bottom:.0001pt"><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif""> <o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Arial","sans-serif"">Fredrik Ek, PhD, </span><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif""><o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Arial","sans-serif"">Researcher, Chemical Biology & Therapeutics</span><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif""><br>
</span><span style="font-size:10.0pt;font-family:"Arial","sans-serif"">Department of Experimental Medical Science<span class="apple-converted-space"> </span></span><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif""><br>
</span><span style="font-size:10.0pt;font-family:"Arial","sans-serif"">Lund University, BMC D10<span class="apple-converted-space"> </span></span><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif""><br>
</span><span style="font-size:10.0pt;font-family:"Arial","sans-serif"">S-221 84 Lund, SWEDEN<span class="apple-converted-space"> </span></span><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif""><br>
</span><span style="font-size:10.0pt;font-family:"Arial","sans-serif"">------------------------<span class="apple-converted-space"> </span></span><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif""><br>
<a href="mailto:Fredrik.Ek@med.lu.se"><span style="font-family:"Arial","sans-serif"">Fredrik.Ek@med.lu.se</span></a></span><span class="apple-converted-space"><span style="font-size:10.0pt;font-family:"Arial","sans-serif""> </span></span><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif""><br>
</span><span style="font-size:10.0pt;font-family:"Arial","sans-serif"">Phone: +46-727 17 22 05 </span><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif""><o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:13.5pt;font-family:"Helvetica","sans-serif"">_______________________________________________<br>
OpenSPIM mailing list<br>
<a href="mailto:OpenSPIM@openspim.org">OpenSPIM@openspim.org</a><br>
<a href="http://openspim.org/mailman/listinfo/openspim">http://openspim.org/mailman/listinfo/openspim</a><o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:13.5pt;font-family:"Helvetica","sans-serif";color:black">_____________<br>
<br>
Michael Weber<br>
Postdoc, Huisken lab<br>
Max Planck Institute of Molecular Cell Biology and Genetics<br>
Pfotenhauerstrasse 108, 01307 Dresden<br>
Tel. 0049 351/2102837<br>
<br>
<a href="http://www.mpi-cbg.de/huisken">http://www.mpi-cbg.de/huisken</a><o:p></o:p></span></p>
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