@article{oai:nagoya.repo.nii.ac.jp:00011948,
 author = {Krone, Annette and Wittbrodt, Joachim},
 journal = {The Fish Biology Journal Medaka},
 month = {},
 note = {Cryoconservation of sperm is a very suitable strategy for the preservation of important strains or mutants. Although this procedure is routinely used in aqua culture for a variety of different species (Rana and Me Andrew, 1989, Steyn and van Vuren, 1987, Wheeler and Thorgaard, 1991), an efficient protocol that allow reliable sperm cryoconservation of small freshwater fish without a need to kill precious males are hardly found. Poorly defined media like milk powder, egg extracts or fetal calf serum used as cryoprotectants (Aoki et al., 1997, Chao et al., 1987, Wheeler and Thorgaard, 1991) may be the cause for variable fertilization rates. Here we describe a rapid and reliable protocol that uses sucrose and DMSO as cryoprotectants (Holtz, 1993). Our protocol allows to efficiently freeze and recover sperm obtained from living fish. In brief sperm is obtained from the males by gently squeezing their testis with a pair of forceps. The sperm is subsequently mixed with freezing medium in a glass capillary. Sperm containing capillaries are brought to successively lower temperatures and are eventually stored in liquid nitrogen. Successful recovery of the preserved stock after several month of storage in liquid nitrogen is routinely achieved by in vitro fertilization., Technical Report},
 pages = {47--48},
 title = {A simple and reliable protocol for cryopreservation of Medaka (Oryzias latipes) spermatozoa},
 volume = {9},
 year = {1997}
}