<html><head><meta http-equiv="Content-Type" content="text/html charset=iso-8859-1"><base href="x-msg://6/"></head><body style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space; ">Hey Fredrik,<div><br></div><div>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.</div><div><br></div><div>Best,</div><div>Michael</div><div><br></div><div><br><div><div>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:</div><br class="Apple-interchange-newline"><blockquote type="cite"><div fpstyle="1" ocsi="0" style="font-family: Helvetica; font-size: medium; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: normal; orphans: 2; text-align: -webkit-auto; text-indent: 0px; text-transform: none; white-space: normal; widows: 2; word-spacing: 0px; -webkit-text-size-adjust: auto; -webkit-text-stroke-width: 0px; "><div style="direction: ltr; font-family: Tahoma; font-size: 10pt; "><div style="margin-bottom: 0px; margin-top: 0px; ">Hi all,</div><p style="margin-bottom: 0px; margin-top: 0px; "> </p><div style="margin-bottom: 0px; margin-top: 0px; ">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.</div><p style="margin-bottom: 0px; margin-top: 0px; "> </p><div style="margin-bottom: 0px; margin-top: 0px; ">Thanks!</div><p style="margin-bottom: 0px; margin-top: 0px; "> </p><div style="margin-bottom: 0px; margin-top: 0px; ">Fredrik</div><div><p style="margin-bottom: 0px; margin-top: 0px; "> </p><div style="font-size: 13px; font-family: Tahoma; "><div style="margin-bottom: 0px; margin-top: 0px; "><font size="2" face="Arial">Fredrik Ek, PhD, </font></div><div style="margin-bottom: 0px; margin-top: 0px; "><font size="2"><font face="Arial">Researcher, Chemical Biology & Therapeutics</font><br><font face="Arial">Department of Experimental Medical Science<span class="Apple-converted-space"> </span></font><br><font face="Arial">Lund University, BMC D10<span class="Apple-converted-space"> </span></font><br><font face="Arial">S-221 84 Lund, SWEDEN<span class="Apple-converted-space"> </span></font><br><font face="Arial">------------------------<span class="Apple-converted-space"> </span></font><br></font><a href="mailto:Fredrik.Ek@med.lu.se"><font size="2" face="Arial">Fredrik.Ek@med.lu.se</font></a><font size="2"><font face="Arial"><span class="Apple-converted-space"> </span></font><br><font face="Arial">Phone: +46-727 17 22 05 </font><br></font></div></div></div></div>_______________________________________________<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><br></div></blockquote></div><br><div apple-content-edited="true">
<span class="Apple-style-span" style="border-collapse: separate; border-spacing: 0px; "><div style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space; "><span class="Apple-style-span" style="border-collapse: separate; color: rgb(0, 0, 0); font-family: Helvetica; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: normal; orphans: 2; text-align: -webkit-auto; text-indent: 0px; text-transform: none; white-space: normal; widows: 2; word-spacing: 0px; border-spacing: 0px; -webkit-text-decorations-in-effect: none; -webkit-text-size-adjust: auto; -webkit-text-stroke-width: 0px; font-size: medium; "><div style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space; "><span class="Apple-style-span" style="border-collapse: separate; color: rgb(0, 0, 0); font-family: Helvetica; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: normal; orphans: 2; text-align: -webkit-auto; text-indent: 0px; text-transform: none; white-space: normal; widows: 2; word-spacing: 0px; border-spacing: 0px; -webkit-text-decorations-in-effect: none; -webkit-text-size-adjust: auto; -webkit-text-stroke-width: 0px; font-size: medium; "><div style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space; ">_____________<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></div></span></div></span></div></span>
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