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A high-throughput method for zebrafish sperm cryopreservation and in vitro fertilization.

Draper, Bruce W and Moens, Cecilia B (2009) A high-throughput method for zebrafish sperm cryopreservation and in vitro fertilization. Journal of visualized experiments : JoVE (29). p. 1395. ISSN 1940-087X

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Article URL: http://www.jove.com/index/Details.stp?ID=1395

Abstract

This is a method for zebrafish sperm cryopreservation that is an adaptation of the Harvey method (Harvey et al., 1982). We have introduced two changes to the original protocol that both streamline the procedure and increase sample uniformity. First, we normalize all sperm volumes using freezing media that does not contain the cryoprotectant. Second, cryopreserved sperm are stored in cryovials instead of capillary tubes. The rates of sperm freezing and thawing (delta degrees C/time) are probably the two most critical variables to control in this procedure. For this reason, do not substitute different tubes for those specified. Working in teams of 2 it is possible to freeze the sperm of 100 males per team in approximately 2 hrs. Sperm cryopreserved using this protocol has an average of 25% fertility (measured as the number of viable embryos generated in an in vitro fertilization divided by the total number of eggs fertilized) and this percent fertility is stable over many years.

Item Type: Article
DOI: 10.3791/1395
PubMed ID: 19581874
NIHMSID: NIHMS137494
PMCID: PMC2785217
Grant Numbers: R01 HG002995
Keywords or MeSH Headings: * Animals * Cryopreservation/methods* * Fertilization in Vitro/methods* * Male * Semen Preservation/methods* * Zebrafish*
Depositing User: Library Staff
Date Deposited: 03 Aug 2009 23:29
Last Modified: 14 Feb 2012 14:42
URI: http://authors.fhcrc.org/id/eprint/323

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