[Zbrafish] Re: Zebrafish sperm cryopreservation

finchg from ohsu.edu via zbrafish%40net.bio.net (by finchg from ohsu.edu)
Fri Oct 17 18:26:56 EST 2008


On Oct 16, 11:08 am, "Kotredes, Kevin"
<Kevin_Kotre... from DFCI.HARVARD.EDU> wrote:
> I'm interested in learning and practicing the procedure to prepare zebrafish
> sperm for cryopreservation and later use for in vitro fertilization.  Does
> anybody have a useful protocol they are willing to share or a resource that they
> can suggest?  Thank you very much!
>
> Kevin P Kotredes
> Research Technician
> Look Zebrafish Lab
> Jimmy Fund Building, RM G03
> Dana-Farber Cancer Institute
> Phone: 617-632-3545
> Email: Kevin_Kotre... from dfci.harvard.edu
>
> The information transmitted in this electronic communication is intended only
> for the person or entity to whom it is addressed and may contain confidential
> and/or privileged material. Any review, retransmission, dissemination or other
> use of or taking of any action in reliance upon this information by persons or
> entities other than the intended recipient is prohibited. If you received this
> information in error, please contact the Compliance HelpLine at 800-856-1983 and
> properly dispose of this information.

Kevin,
I'm trying to start up sperm cryopreservation at my lab as well.
Multiple sperm freeze protocols can be found at the ZIRC site:
http://zebrafish.org/zirc/documents/protocols.php
Also, there is a protocol in the Christiane Nusslein-Volhard/Ralf Dahm
Zfish book:
http://books.google.com/books?id=zS-5CKOjCIgC&pg=PA30&dq=zebrafish+sperm+cryopreservation
The Draper protocol appears to be the most widely used (e.g. at ZIRC,
in TILLING at Moens Lab, etc.) and is well written/easy to follow.

....however, I am following it and thus far have been having a really
difficult time getting the 1-3ul of sperm that the protocol
recommends.  If anyone reading this can recommend or describe a
squeezing technique that can produce larger volumes, please please
take the time to post some advice.  I have tried everything I can
think of, variety of forceps, fingers, variety of pressures, variety
of anatomical regions, sexy music, etc.  Anyone?  Please........



More information about the Zbrafish mailing list