Hello Di Jiang,
First, wash, wash, and wash your eggs again, making sure
there is no dissected tissue (enzymes, bacteria etc.) mixed in with
them following dissection. Second, set up a series of petri dishes,
each with a bunch of eggs, and fertilize them with fresh sperm. Wash
the first batch after 1 min following fertilization, the second batch
after 2 min following fertilization, the 3rd batch after 4 min
following fertilization, and the fourth batch after 8 min following
fertilization. One of these treatments will almost certainly give
you higher fert. success than the others. All of this should be done
in filtered seawater.
A second replication of this experiment should be done in
filtered AND pasteurized seawater (heat it too 80 C for 15 min and do
not let it boil!). You also might also try a third replication in
Jamarin-U (artificial seawater). There is something in Jam-U (trade
secret) that some embryos like and it is not uncommon for them to
fertilize and develop better than in filtered, or filtered/
pasteurized seawater.
Good luck!
Chris
>Hi list members:
>
>I am trying to get synchronized embryos at different stages (early,
>mid, and late tail
>bud stages, and after hatching) from Styela plicata and Styela clava
>by dissection
>followed by cross fertilization. I have been able to get a lot of
>gametes but the
>fertilization is very low. I grow them at 18 C in a 15 cm Petri
>dish with sperm and
>eggs together including the dissected tissues. Does anyone know
>some tricks to
>increase the fertilization rate?
>
>Di Jiang
>UCSB
--
Chris Cameron, NSERC Post-Doctoral Fellow
*-----------------------------------------------------------------*
Department of Biology telephone: (250) 592-2649
University of Victoria fax: (250) 721-7120
Victoria, BC, Canada, V8W 2Y2
*-----------------------------------------------------------------*
http://cluster3.biosci.utexas.edu/faculty/cameronc/CBC.htm
(research interests, publications, Hemichordata)
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