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

Bruce W. Draper, Cecilia B. Moens

Research output: Contribution to journalArticle

16 Citations (Scopus)

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 (δ°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 ~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.

Original languageEnglish (US)
Article numbere1395
JournalJournal of Visualized Experiments
Issue number29
DOIs
StatePublished - Jul 2009

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Cryopreservation
Zebrafish
Fertilization in Vitro
Freezing
Spermatozoa
Throughput
Capillary tubes
Thawing
Fertility
Zygote
Embryonic Structures

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

Cite this

A high-throughput method for zebrafish sperm cryopreservation and in vitro fertilization. / Draper, Bruce W.; Moens, Cecilia B.

In: Journal of Visualized Experiments, No. 29, e1395, 07.2009.

Research output: Contribution to journalArticle

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