[Zbrafish] Re: Imaging Zebrafish

[Roy Edward]

On May 22, 8:27 am, FritzXC23 <frits... from gettysburg.edu> wrote:
> Hi All!
>
> My name is Sarah Fritz and I am undergraduate student from Gettysburg
> College. I am currently working on a research project that involves
> imaging fluorescent zebrafish hair cells. I am wondering if anyone has
> suggestions about the best way to get 5 dpf fish to lay on their sides
> without having to mount them. Thanks!
>
> Sarah Fritz

Hi Sarah

Excuse the commercial response but we have developed a technology that
should help in this respect.
CyGEL/CyGEL Sustain are thermoreversible hydrogels that are a liquid
when cooled and gel when warmed above room temperature.
These were initially developed to allow fluorescent imaging of live
non-adherent cells, but in collaboration with a zebrafish lab have
proved very useful for zebrafish embryos.

For the live embryo work we have found CyGEL Sustain to be best.
It is optically inert and clear.  It accepts embryo medium (40-60X is
best to avoid over-dilution of the gel) and MS-222 and analogues.  We
have added MS-222 at a 2X final conc. compared to that used to
initially anaesthetize the embryos, creating a reservoir of
anaesthetic for the period of the procedure.
Embryos can be moved into position by melting the gel, moving them and
re-setting the gel.
It can be used on slides with coverslips or in microplate wells.
This process can be used for imaging or for micro-injection.  In all
cases embryos MUST be de-chorionated.
Embryos can be recovered, live, after an hour in the gel, requiring
only a gentle wash to remove residual gel and time to allow them to re-
awaken!

CyGEL (a PBS-based equivalent) is more suitable for fixed embryo
imaging when special medium is not required, and is provided with 40X
PBS.

A poster describing CyGEL Sustain's use for live embryo manipulation
will be presented by collaborators at the forthcoming zebrafish
conference in Rome.
I have also collaborated with C elegans and parasite researcher
teams.  Again we were able successfully to image and recover live
organisms.
Full details on CyGEL Sustain can be found at our website.  I can also
provide a copy of a previous poster I presented at SBS on our CyGEL
technology.

Very best regards
Roy

Roy Edward
roy from biostatus.com

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