PUBLICATION
A high-throughput method for zebrafish sperm cryopreservation and in vitro fertilization
- Authors
- Draper, B.W., and Moens, C.B.
- ID
- ZDB-PUB-090714-13
- Date
- 2009
- Source
- Journal of visualized experiments : JoVE (29): (Journal)
- Registered Authors
- Draper, Bruce, Moens, Cecilia
- Keywords
- none
- MeSH Terms
-
- Animals
- Cryopreservation/methods*
- Fertilization in Vitro/methods*
- Male
- Semen Preservation/methods*
- Zebrafish*
- PubMed
- 19581874 Full text @ J. Vis. Exp.
Citation
Draper, B.W., and Moens, C.B. (2009) A high-throughput method for zebrafish sperm cryopreservation and in vitro fertilization. Journal of visualized experiments : JoVE. (29).
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping