[Xenopus] MISC

Thu Oct 29 09:01:41 EDT 2020

Hi Anna, animal caps are a piece of cake compared to isolating endoderm. If you’re like my undergrads, though, and over proteinase K treat your fixed embryos, the ectoderm comes right off however!!  This may not be ideal for an intended isolation of endoderm, but a gentle protease (or better, collagenase) might work if carefully controlled.   Best, Rob

From: <xenopus-bounces at lists.mbl.edu> on behalf of Anna Philpott <ap113 at cam.ac.uk>
Date: Thursday, October 29, 2020 at 8:57 AM
To: "Green, Jeremy" <jeremy.green at kcl.ac.uk>, Leon Peshkin <peshkin at gmail.com>
Cc: "xenopus at lists.mbl.edu" <xenopus at lists.mbl.edu>
Subject: Re: [Xenopus] MISC

Dear Leon,
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?
Anna

Prof. Anna Philpott
Head of the School of Biological Sciences
And
Group Leader, Cambridge Stem Cell Institute
University of Cambridge

From: <xenopus-bounces at lists.mbl.edu> on behalf of Jeremy Green <jeremy.green at kcl.ac.uk>
Date: Thursday, 29 October 2020 at 11:13
To: Leon Peshkin <peshkin at gmail.com>
Cc: "xenopus at lists.mbl.edu" <xenopus at lists.mbl.edu>
Subject: Re: [Xenopus] MISC

Dear Leon,

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.

Best wishes,

Jeremy

Jeremy B.A. Green Ph.D.
Vice-Dean International (Research)
Professor of Developmental Biology
Centre for Craniofacial & Regenerative Biology
King's College London
Guy's Tower, Floor 27
London SE1 9RT
UK

jeremy.green at kcl.ac.uk<mailto:jeremy.green at kcl.ac.uk>
Tel. +44 20 7188 1794
https://www.kcl.ac.uk/people/jeremy-green

On 29 Oct 2020, at 03:45, Leon Peshkin <peshkin at gmail.com<mailto:peshkin at gmail.com>> wrote:

Dear Colleagues !

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 !

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.
   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.

Does someone have a protocol and experience for blood perfusion in Xenopus ?

   THANK YOU

   - L. Peshkin

_______________________________________________
Xenopus mailing list
Xenopus at lists.mbl.edu<mailto:Xenopus at lists.mbl.edu>
https://eur03.safelinks.protection.outlook.com/?url=https%3A%2F%2Flists.mbl.edu%2Fmailman%2Flistinfo%2Fxenopus&amp;data=04%7C01%7Cjeremy.green%40kcl.ac.uk%7Cf73f268fc9bc43ab49ba08d87bbed42e%7C8370cf1416f34c16b83c724071654356%7C0%7C0%7C637395406857832409%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=4Vaj%2BbdjlhcF2eH0mcUjy9tAwg7NJTbYkVPYzduZocw%3D&amp;reserved=0

-------------- next part --------------
An HTML attachment was scrubbed...
URL: http://lists.mbl.edu/pipermail/xenopus/attachments/20201029/12df03d8/attachment-0001.html 