[Zbrafish] Re: In vitro Fertilization

karen.leewingman from gmail.com via zbrafish%40net.bio.net (by karen.leewingman from gmail.com)
Tue Mar 11 03:14:33 EST 2008


Salut,

For the oocytes collection, they do not give oocytes everyday.  They
do have a cycle.  So usually there are only a few female fish have
good quality oocytes per day.  Try to squeeze the female within an
hour after the light comes on.  The oocytes will be re-absorbed and
there are no good for fertilization anymore.  For the sperm, it is
fairly easy to collect.  When you squeeze the male, the whitish liquid
is the sperm already.  To check the motility of the sperm, you can put
a drop (you collect the sperm and put them into the sperm extender
solution) of the sperm onto the slide and use a 10X objective to have
a look.  It is not motile, but as soon as you add some water onto it,
the sperm will start to swim. For more information, please check the
following paper.
Lee, K.W., Webb, S.E., and Miller, A.L. (1999). A wave of free
cytosolic calcium traverses zebrafish eggs on activation.
Developmental Biology 214: 168-180.

If you need any further information, please feel free to contact me at
karen.leewingman from gmail.com (Paris).



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