Feasibility of vitrification of zebrafish (Danio rerio) embryos using methanol

Authors: Liu, X.-H., Zhang, T. and Rawson, D.M.

Journal: CryoLetters

Volume: 19

Pages: 309-318

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Authors: Liu, X.H., Zhang, T. and Rawson, D.M.

Journal: Cryo-Letters

Volume: 19

Issue: 5

Pages: 309-318

ISSN: 0143-2044

This study investigated the feasibility of vitrification of zebrafish embryos using methanol as the cryoprotectant. The lowest apparent vitrification concentration of methanol was found to be 10M for 1μl volume. The toxicity of 10M methanol was much more pronounced with early stage (1-cell and 64-cell) embryos than later stages (50% epiboly, 6-somite and prim-6). Embryos at both early stages withstood 3min exposure to 10M methanol at 0°C. However, the maximum time periods the 50% epiboly, 6-somite and prim-6 stage embryos could be exposed to 10M methanol without significant detrimental effects were 10, 5, 10min respectively. No embryo survived after 40min exposure. Vitrification of embryos on gold electron microscope grids showed higher percentage of embryos remaining morphologically intact, ~50% and 80% for early and late stage embryos respectively, compared with previous studies. However, no embryos showed any developmental ability.

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