Cryopreservation method for Drosophila melanogaster embryos

Li Zhan; Min gang Li; Thomas Hays; John Bischof

doi:10.1038/s41467-021-22694-z

Cryopreservation method for Drosophila melanogaster embryos

Li Zhan, Min gang Li, Thomas Hays, John Bischof

Research output: Contribution to journal › Article › peer-review

35 Scopus citations

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.

Original language	English (US)
Article number	2412
Journal	Nature communications
Volume	12
Issue number	1
DOIs	https://doi.org/10.1038/s41467-021-22694-z
State	Published - Dec 1 2021

Bibliographical note

Funding Information:
We thank Dr. Amanda Neisch for productive discussions surrounding the genetics and variation in cryopreservation. We thank Zonghu Han and Anthony Chen for serving as volunteers to test the robustness of the developed cryopreservation protocol. We thank Dr. Michael Etheridge for his critical reading and comments to the manuscript. Funding from the US National Institutes of Health (NIH) (1R21OD028758) is gratefully acknowledged. T.H. further acknowledges NIH GM R01GM044757 and J.B. acknowledges NSF EEC 1941543. L.Z. acknowledges the support of a Doctoral Dissertation Fellowship at the University of Minnesota.

Publisher Copyright:
© 2021, The Author(s).

PubMed: MeSH publication types

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

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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.",

author = "Li Zhan and Li, {Min gang} and Thomas Hays and John Bischof",

note = "Funding Information: We thank Dr. Amanda Neisch for productive discussions surrounding the genetics and variation in cryopreservation. We thank Zonghu Han and Anthony Chen for serving as volunteers to test the robustness of the developed cryopreservation protocol. We thank Dr. Michael Etheridge for his critical reading and comments to the manuscript. Funding from the US National Institutes of Health (NIH) (1R21OD028758) is gratefully acknowledged. T.H. further acknowledges NIH GM R01GM044757 and J.B. acknowledges NSF EEC 1941543. L.Z. acknowledges the support of a Doctoral Dissertation Fellowship at the University of Minnesota. Publisher Copyright: {\textcopyright} 2021, The Author(s).",

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N2 - 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.

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Cryopreservation method for Drosophila melanogaster embryos

Abstract

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ATP-Bio: NSF Engineering Research Center for Advanced Technologies for the Preservation of Biological Systems (ATP-Bio)

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Cryopreservation method for Drosophila melanogaster embryos

Abstract

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ATP-Bio: NSF Engineering Research Center for Advanced Technologies for the Preservation of Biological Systems (ATP-Bio)

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