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Cryopreservation method for Drosophila melanogaster embryos

Author

Listed:
  • Li Zhan

    (University of Minnesota
    University of Minnesota)

  • Min-gang Li

    (University of Minnesota)

  • Thomas Hays

    (University of Minnesota)

  • John Bischof

    (University of Minnesota
    University of Minnesota
    University of Minnesota)

Abstract

The development of a widely adopted cryopreservation method remains a major challenge in Drosophila research. Here we report a robust and easily implemented cryopreservation protocol of Drosophila melanogaster embryos. We present innovations for embryo permeabilization, cryoprotectant agent loading, and rewarming. We show that the protocol is broadly applicable, successfully implemented in 25 distinct strains from different sources. We demonstrate that for most strains, >50% embryos hatch and >25% of the resulting larvae develop into adults after cryopreservation. We determine that survival can be significantly improved by outcrossing to mitigate the effect of genetic background for strains with low survival after cryopreservation. We show that flies retain normal sex ratio, fertility, and original mutation after successive cryopreservation of 5 generations and 6-month storage in liquid nitrogen. Lastly, we find that non-specialists are able to use this protocol to obtain consistent results, demonstrating potential for wide adoption.

Suggested Citation

  • Li Zhan & Min-gang Li & Thomas Hays & John Bischof, 2021. "Cryopreservation method for Drosophila melanogaster embryos," Nature Communications, Nature, vol. 12(1), pages 1-10, December.
    • Handle: RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-22694-z
    DOI: 10.1038/s41467-021-22694-z
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    Cited by:

    1. Li Zhan & Zonghu Han & Qi Shao & Michael L. Etheridge & Thomas Hays & John C. Bischof, 2022. "Rapid joule heating improves vitrification based cryopreservation," Nature Communications, Nature, vol. 13(1), pages 1-15, December.
    2. Matthew J. Powell-Palm & E. Michael Henley & Anthony N. Consiglio & Claire Lager & Brooke Chang & Riley Perry & Kendall Fitzgerald & Jonathan Daly & Boris Rubinsky & Mary Hagedorn, 2023. "Cryopreservation and revival of Hawaiian stony corals using isochoric vitrification," Nature Communications, Nature, vol. 14(1), pages 1-11, December.

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