PUBLICATION

Cryopreservation and Laser Nanowarming of Zebrafish Embryos Followed by Hatching and Spawning

Authors
Khosla, K., Kangas, J., Liu, Y., Zhan, L., Daly, J., Hagedorn, M., Bischof, J.
ID
ZDB-PUB-201002-216
Date
2020
Source
Advanced biosystems   4(11): e2000138 (Journal)
Registered Authors
Hagedorn, Mary
Keywords
gold nanoparticles, photothermal heating, vitrification, zebrafish
MeSH Terms
  • Animals
  • Cryopreservation/methods*
  • Cryoprotective Agents
  • Embryo Culture Techniques/methods*
  • Embryo, Nonmammalian/physiology*
  • Gold/chemistry
  • Lasers
  • Metal Nanoparticles/chemistry
  • Nanotechnology/methods*
  • Nanotubes/chemistry
  • Vitrification
  • Zebrafish
PubMed
32996298 Full text @ Adv Biosyst
Abstract
This study shows for the first time the ability to rewarm cryopreserved zebrafish embryos that grow into adult fish capable of breeding normally. The protocol employs a single injection of cryoprotective agents (CPAs) and gold nanorods (GNRs) into the yolk and immersion in a precooling bath to dehydrate the perivitelline space. Then embryos are encapsulated within CPA and GNR droplets, plunged into liquid nitrogen, cryogenically stabilized, and rewarmed by a laser pulse. Postlaser nanowarming, embryos (n = 282) exhibit intact structure by 1 h (40%), continued development after 3 h (22%), movement after 24 h (11%), hatching after 48 h (9%), and swimming after Day 5 (3%). Finally, from fish that survives till Day 5, two larvae are grown to adulthood and spawned, yielding survival comparable to an unfrozen control. Future efforts will focus on improving the survival to adulthood and developing methods to cryopreserve large numbers of embryos for research, aquaculture, and biodiversity preservation.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping