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preserve FISH sperm.

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preserve FISH sperm.

Postby blueberry94 » Sat Mar 12, 2011 1:43 pm

Hello, my name is Oliver,and i love biologie :mrgreen: ,but i need some help if possible.

Last summer i bought a couple of fish that cost me 100$ and unfortunately i've lost the male yesterday morning.. :(
The female is in great health.. But now the male is dead,so i can't breed them anymore..
So I've thinked is it possible after extracting the male testicles,(i know their place,so by dissecting the male i could remove his testicles,and fertilize the eggs of the female). But the problem is not really here...
The sperm will die in few hours.. so i was asking if you have any thing to do to conserve the sperm for a little more time.. Because the female is now in condition to breed with another fish.. i will let her ,by exitation,to drop her eggs(it is possible with the fish that i'm breeding)that are surely not fertilized and fertilize them with the sperm that i'll collect from the dead male....
The real probleme is,how to conserve the sperm for a few days? because as i know it won't live so much(~48-72h) after the death of the fish.. :(

Thanks a lot!!
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Postby blueberry94 » Sat Mar 12, 2011 3:55 pm

What about the formol(formalin)?? can i use it?? to concerve the sperm. while i got the eggs to fertilize them?
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Postby JackBean » Sat Mar 12, 2011 5:23 pm

definitely not formalin! That will form bridges between proteins and thus kill them.

You could try to add something like glycerol (used to freeze bacteria or yeasts and molecules), sucrose (used for cryopreservation of some plant tissues) and freeze, best at -80°C, but I guess you do not have such freezer :lol: so -20°C should be fine
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
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Postby blueberry94 » Sat Mar 12, 2011 5:43 pm

Hello,
Thanks for you answer :)
of i will buy glycerol from the pharmacy,(should i buy also sucrose?? i'm working with animals not plants,so i think i don't need sucrose)

One more question please..
I will dissect the testicles of the male,can i just put them in the glycerol solution that i bought?? and freez?? or should i do something else??
Please if you could reply quickly because the sperm life won't be for too long,so i must react quickly
Thanks a lot for your help JACKBEAN
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Postby JackBean » Sat Mar 12, 2011 5:49 pm

I just said, I know it's used for plant cryopreservation, but I'm not sure, whether is it used also for animals. I know, you're not working with plants :lol:

It would be best to have some buffer, but I guess you won't get any, so I would recommend at least some isotonic solution (maybe something like for the eye lens?) and from that volume, the glycerol should be about 20%.
(I don't know, how much will you need, but let's say you will have 4 ml of the solution and 1 ml of the glycerol)
http://www.biolib.cz/en/main/

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Postby blueberry94 » Sat Mar 12, 2011 6:06 pm

mmm.. can i use sterimar?? it is isotonic:

STÉRIMAR is made from pure sea water. These waters are monitored by an independent regulatory authority to ensure sanitary standards. It is then:

- filtered and sterilised
====>{ diluted to 1/3, which makes it isotonic (appropriate for the human body) }
Sea water has the added benefit of containing various naturally occurring trace elements, many of which have been shown to have antiseptic (silver & zinc), anti-inflammatory (copper) or anti-allergic (manganese) properties.


Well as i understand i just have to dissect the testicles, and put them in the STERIMAR solution which contain glycerol solution and in the fredge? it's all ?
Thanks a lot :) if i succeed to breed i will share some photos :)
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Postby JackBean » Sat Mar 12, 2011 6:14 pm

yeah, that could work. Although I really have NO idea, what will the sperm do in only-isotonic solution with glycerol at -20°C
http://www.biolib.cz/en/main/

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Postby blueberry94 » Sun Mar 13, 2011 7:42 am

Thanks a lot!! i hope it will give me 48 more hours...

I hope!
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