<div dir="ltr"><div dir="ltr">Hi Anna et al.<div>In my day there was no Gastromaster and we cut loads of animal caps with sharpened tungsten needles and a hair loop or forceps to steady the embryo. To separate layers in neurulae it helps a lot to include a little trypsin in the medium. Leave the Ca in then you tend to degrade the ECM but not to disaggregate the cells. Methods are in the attached (otherwise uninteresting) paper.</div><div>Good luck,</div><div>Jonathan</div><div><br></div><div><p style="margin-top:0px;margin-bottom:0px;font-family:Calibri,Arial,Helvetica,sans-serif">Jonathan M.W.Slack, PhD, FMedSci.,</p><p style="margin-top:0px;margin-bottom:0px;font-family:Calibri,Arial,Helvetica,sans-serif">Emeritus Professor, University of Bath, UK.</p><p style="margin-top:0px;margin-bottom:0px;font-family:Calibri,Arial,Helvetica,sans-serif">Emeritus Professor, University of Minnesota, USA.</p><p style="margin-top:0px;margin-bottom:0px"><a href="http://www.bath.ac.uk/bio-sci/contacts/academics/jonathan_slack/" target="_blank">http://www.bath.ac.uk/bio-sci/contacts/academics/jonathan_slack/</a></p></div></div></div><br><div class="gmail_quote"><div dir="ltr" class="gmail_attr">On Thu, Oct 29, 2020 at 1:27 PM Klein, Peter <<a href="mailto:pklein@pennmedicine.upenn.edu" target="_blank">pklein@pennmedicine.upenn.edu</a>> wrote:<br></div><blockquote class="gmail_quote" style="margin:0px 0px 0px 0.8ex;border-left:1px solid rgb(204,204,204);padding-left:1ex">
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<p class="MsoNormal">Hi Leon, <u></u><u></u></p>
<p class="MsoNormal"><u></u> <u></u></p>
<p class="MsoNormal">The gastromaster is much faster but in my lab only works with the younger generation of scientists, who could easily cut 50 or more in an hour. Either approach takes a little practice, but given shortage of tips and lack of information
about how to make them, I agree with above: you could cut hundreds with a good team, some #5 Dumonts, and hair knives.<u></u><u></u></p>
<p class="MsoNormal"><u></u> <u></u></p>
<p class="MsoNormal">Peter<u></u><u></u></p>
<p class="MsoNormal"><u></u> <u></u></p>
<p class="MsoNormal"><u></u> <u></u></p>
<div style="border-right:none;border-bottom:none;border-left:none;border-top:1pt solid rgb(181,196,223);padding:3pt 0in 0in">
<p class="MsoNormal"><b><span style="font-size:12pt;color:black">From: </span></b><span style="font-size:12pt;color:black"><<a href="mailto:xenopus-bounces@lists.mbl.edu" target="_blank">xenopus-bounces@lists.mbl.edu</a>> on behalf of Anna Philpott <<a href="mailto:ap113@cam.ac.uk" target="_blank">ap113@cam.ac.uk</a>><br>
<b>Date: </b>Thursday, October 29, 2020 at 8:48 AM<br>
<b>To: </b>"Green, Jeremy" <<a href="mailto:jeremy.green@kcl.ac.uk" target="_blank">jeremy.green@kcl.ac.uk</a>>, Leon Peshkin <<a href="mailto:peshkin@gmail.com" target="_blank">peshkin@gmail.com</a>><br>
<b>Cc: </b>"<a href="mailto:xenopus@lists.mbl.edu" target="_blank">xenopus@lists.mbl.edu</a>" <<a href="mailto:xenopus@lists.mbl.edu" target="_blank">xenopus@lists.mbl.edu</a>><br>
<b>Subject: </b>[External] Re: [Xenopus] MISC<u></u><u></u></span></p>
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<div>
<p class="MsoNormal"><u></u> <u></u></p>
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<p class="MsoNormal"><span>Dear Leon,</span><u></u><u></u></p>
<p class="MsoNormal"><span>OMG I would LOVE a cap cutting jamboree. We have a gastromaster on the shelf with some tips but also found it less than the hype and as we can’t get tips any more and they break a lot, it’s not
much of a solution. Yes, 1 also think No5s and some experience are the only way forward-freezing in batches if necessary for pooling later. Do you need caps rather than animal poles? Razor blades are quicker obvs. We are currently trying to compare ecto/meso
with endoderm. This unfortunately means peeling the epidermis off the ventral tissue (neurula stages) to get pure endoderm. This is agonising. Does anyone have an easy way of doing that?</span><u></u><u></u></p>
<p class="MsoNormal"><span>Anna</span><u></u><u></u></p>
<p class="MsoNormal"><span> </span><u></u><u></u></p>
<p class="MsoNormal"><span> </span><u></u><u></u></p>
<p class="MsoNormal"><span> </span><u></u><u></u></p>
<div>
<p class="MsoNormal"> <u></u><u></u></p>
<p class="MsoNormal"> <u></u><u></u></p>
<p class="MsoNormal">Prof. Anna Philpott<u></u><u></u></p>
<p class="MsoNormal">Head of the School of Biological Sciences<u></u><u></u></p>
<p class="MsoNormal">And<u></u><u></u></p>
<p class="MsoNormal">Group Leader, Cambridge Stem Cell Institute<u></u><u></u></p>
<p class="MsoNormal">University of Cambridge<u></u><u></u></p>
<p class="MsoNormal"> <u></u><u></u></p>
</div>
<p class="MsoNormal"><span> </span><u></u><u></u></p>
<p class="MsoNormal"><span> </span><u></u><u></u></p>
<div style="border-right:none;border-bottom:none;border-left:none;border-top:1pt solid rgb(181,196,223);padding:3pt 0in 0in">
<p class="MsoNormal"><b><span style="font-size:12pt;color:black">From: </span></b><span style="font-size:12pt;color:black"><<a href="mailto:xenopus-bounces@lists.mbl.edu" target="_blank">xenopus-bounces@lists.mbl.edu</a>> on behalf of Jeremy Green <<a href="mailto:jeremy.green@kcl.ac.uk" target="_blank">jeremy.green@kcl.ac.uk</a>><br>
<b>Date: </b>Thursday, 29 October 2020 at 11:13<br>
<b>To: </b>Leon Peshkin <<a href="mailto:peshkin@gmail.com" target="_blank">peshkin@gmail.com</a>><br>
<b>Cc: </b>"<a href="mailto:xenopus@lists.mbl.edu" target="_blank">xenopus@lists.mbl.edu</a>" <<a href="mailto:xenopus@lists.mbl.edu" target="_blank">xenopus@lists.mbl.edu</a>><br>
<b>Subject: </b>Re: [Xenopus] MISC</span><u></u><u></u></p>
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<div>
<p class="MsoNormal"> <u></u><u></u></p>
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<p class="MsoNormal">Dear Leon, <u></u><u></u></p>
<div>
<p class="MsoNormal"> <u></u><u></u></p>
</div>
<div>
<p class="MsoNormal">With a bit of practice and well-sharpened Dumont #5 forceps, a person can cut around 200 caps in an hour. I suggest that if you want to get many hundreds of caps that you simply have a cap-cutting jamboree - not so much biting the bullet
as having a party. Seriously. I think you’ll have no problem getting volunteers and (COVID, etc. permitting), I’d be happy to come over myself.<u></u><u></u></p>
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<div>
<p class="MsoNormal"> <u></u><u></u></p>
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<p class="MsoNormal">Best wishes,<u></u><u></u></p>
</div>
<div>
<p class="MsoNormal"> <u></u><u></u></p>
</div>
<div>
<p class="MsoNormal">Jeremy<u></u><u></u></p>
</div>
<div>
<p class="MsoNormal"> <u></u><u></u></p>
<div>
<div>
<div>
<div>
<div>
<div>
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<p class="MsoNormal"><span style="font-family:Helvetica;color:black">Jeremy B.A. Green Ph.D.</span><u></u><u></u></p>
</div>
<div>
<p class="MsoNormal"><span style="font-family:Helvetica;color:black">Vice-Dean International (Research)</span><u></u><u></u></p>
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<p class="MsoNormal"><span style="font-family:Helvetica;color:black">Professor of Developmental Biology</span><u></u><u></u></p>
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<p class="MsoNormal"><span style="font-family:Helvetica;color:black">Centre for Craniofacial & Regenerative Biology</span><u></u><u></u></p>
</div>
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<p class="MsoNormal"><span style="font-family:Helvetica;color:black">King's College London</span><u></u><u></u></p>
</div>
<div>
<p class="MsoNormal"><span style="font-family:Helvetica;color:black">Guy's Tower, Floor 27</span><u></u><u></u></p>
</div>
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<p class="MsoNormal"><span style="font-family:Helvetica;color:black">London SE1 9RT</span><u></u><u></u></p>
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<p class="MsoNormal"><span style="font-family:Helvetica;color:black">UK</span><u></u><u></u></p>
</div>
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<p class="MsoNormal" style="margin-bottom:12pt"><span style="font-family:Helvetica;color:black"><br>
<a href="mailto:jeremy.green@kcl.ac.uk" target="_blank">jeremy.green@kcl.ac.uk</a><br>
Tel. +44 20 7188 1794<br>
<a href="https://www.kcl.ac.uk/people/jeremy-green" target="_blank">https://www.kcl.ac.uk/people/jeremy-green</a><br>
<br>
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</span><u></u><u></u></p>
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<p class="MsoNormal"><br>
<br>
<br>
<u></u><u></u></p>
<blockquote style="margin-top:5pt;margin-bottom:5pt">
<div>
<p class="MsoNormal">On 29 Oct 2020, at 03:45, Leon Peshkin <<a href="mailto:peshkin@gmail.com" target="_blank">peshkin@gmail.com</a>> wrote:<u></u><u></u></p>
</div>
<p class="MsoNormal"> <u></u><u></u></p>
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<p class="MsoNormal">Dear Colleagues !<u></u><u></u></p>
</div>
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<p class="MsoNormal"> <u></u><u></u></p>
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<div>
<p class="MsoNormal">I have a few general questions and tried posting to Slack "Xenopus Community" without success. People - Slack is a nice way to keep discussions going, please use it !
<u></u><u></u></p>
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<p class="MsoNormal"> <u></u><u></u></p>
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<div>
<p class="MsoNormal">Has someone worked out a way to make animal caps en masse ? I did hear the gastromaster legend and even obtained one with a few tips … but it does not really work in my hands. Any other devices ? Please do not tell me to just bite the
bullet. I'd like to be able to cut many hundreds. <u></u><u></u></p>
</div>
<div>
<p class="MsoNormal"> There are a couple of animal cap instructional videos with forceps but I know many people use other techniques. Would be nice to have more videos.
<u></u><u></u></p>
</div>
<div>
<p class="MsoNormal"> <u></u><u></u></p>
</div>
<div>
<p class="MsoNormal">Does someone have a protocol and experience for blood perfusion in Xenopus ?
<u></u><u></u></p>
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<div>
<p class="MsoNormal"> <u></u><u></u></p>
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<p class="MsoNormal"> THANK YOU <u></u><u></u></p>
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<p class="MsoNormal"> <u></u><u></u></p>
</div>
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<p class="MsoNormal"> - L. Peshkin <u></u><u></u></p>
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<p class="MsoNormal"> <u></u><u></u></p>
</div>
<div>
<p class="MsoNormal"> <u></u><u></u></p>
</div>
</div>
<p class="MsoNormal">_______________________________________________<br>
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