[Xenopus] high-throughput devitellinizing?

Sat Oct 31 12:03:33 EDT 2020

Hi everyone,

From: Enrique Amaya <Enrique.Amaya at manchester.ac.uk>
> The most time consuming part is removing the vitelline membranes

	Yes!! Speaking of which: does anyone have an enzymatic method for this yet?  Maybe recombinant hatching enzyme cocktail or something? Is there a decent way to devitellinize tons of embryos at once?

Best,

Mike Levin

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-----Original Message-----
From: <xenopus-bounces at lists.mbl.edu> on behalf of Enrique Amaya <Enrique.Amaya at manchester.ac.uk>
Date: Thursday, October 29, 2020 at 7:45 AM
To: Jeremy Green <jeremy.green at kcl.ac.uk>
Cc: Leon Peshkin <peshkin at gmail.com>, "xenopus at lists.mbl.edu" <xenopus at lists.mbl.edu>
Subject: Re: [Xenopus] MISC

    Dear Leon,
    The most time consuming part is removing the vitelline membranes. When I was animal cap king decades ago my preferred method was to use an eye brow hair knife and put the de-vitellinised blastula embryos on their side and decapitate them with the eye brow knife… a very rapid method. If working with another, I would suggest having someone removing the vitelline membranes and another person performing the decapitations, to ensure consistency in animal cap removals. One can easily get through hundreds this way. How many do you need?
    Enrique

    ——————————————————————————————
    Professor Enrique Amaya
    Division of Cell Matrix Biology & Regenerative Medicine
    Michael Smith Building
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    Faculty of Biology, Medicine and Health
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    On 29 Oct 2020, at 11:08, Green, Jeremy <jeremy.green at kcl.ac.uk> wrote:

    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
    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> 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 

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