sperm freezing

[Carrie Jones]

You might find some useful information in The Zebrafish Book, 
Chapter 7:  

http://zfin.org/zf_info/zfbook/cont.html#cont7

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Carrie R. Jones
Zebrafish International Resource Center
5274 University of Oregon
Eugene, OR  97403-5274
Phone: (541) 346-6028 ext. 22
Fax: (541) 346-6151
Email: cjones at zfin.org
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May-Su YOU wrote:
> I am Maysu You, now working at IMCB (Institute of Molecular and Cell
>  Biology) in Singapore. We are now working on sperm 
> squeezing,freezing and in vitro fertilization of Zebrafish. We got
> around  90% of IVF successful rate when we used fresh sperm (from
> both  squeezed sperm and dissected testes) . So we moved to freezing
> step.  We tried two different cryoprotectants (methanol and BSMIS:
> Buffered  sperm motality inhibitor solution). After thawing, we
> cann't get any IVF  embryo. Therefore, we doubt about the freezing
> procedure which we  used at the moment. We also tried to observe the
> sperm, but even with  trypan blue, we are not sure wheather the
> sperm are live or dead.  According to the results,I think there are
> some questions to be  answered by whom has experties in this work   
> 
> 1.Maybe our freezing protocal is not totally correct (is there any
> critical  point that we didn't catch?)  
> 
> 2.How can we check the viability of the sperm? (we observed sperm 
> under 400 time magnification)  
> 
> 3. What will be the best way to fertilize eggs with frozen sperm.
> Your  help will give us a big improvement of our work.Thank you in
> advance.   
> 
> Best regards 
> 
> Maysu 
> 
> 
> ************************************************ 
> Dr. May-Su YOU 
> Research Officer 
> Institute of Molecular and Cell Biology, 
> A*STAR building, 1 Research Link, 
> The National University of Singapore, 
> 117604 Singapore 
> Republic of Singapore 
> 
> Tel: (65) 68727092 (lab) 
> Fax: (65) 68727007 
> e-mail: youms at imcb.a-star.edu.sg 
> 
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> TELEPHONE AND FAX 
> NUMBERS*** 
>