Article

Gold Nanorod Induced Warming of Embryos from the Cryogenic State Enhances Viability

Department of Mechanical Engineering, University of Minnesota at Twin Cities, 111 Church Street SE, Minneapolis, Minnesota 55455, United States
Smithsonian Conservation Biology Institute, Smithsonian National Zoological Park, 3001 Connecticut Avenue NW, Washington, D.C. 20008, United States
§ Hawaii Institute of Marine Biology, University of Hawaii, 46-007 Lilipuna Road, Kaneohe, Hawaii 96744, United States
Department of Biomedical Engineering, University of Minnesota at Twin Cities, 312 Church Street SE, Minneapolis, Minnesota 55455, United States
ACS Nano, 2017, 11 (8), pp 7869–7878
DOI: 10.1021/acsnano.7b02216
Publication Date (Web): July 13, 2017
Copyright © 2017 American Chemical Society
*Tel: 612-625-5513. E-mail: bischof@umn.edu.

Abstract

Abstract Image

Zebrafish embryos can attain a stable cryogenic state by microinjection of cryoprotectants followed by rapid cooling, but the massive size of the embryo has consistently led to failure during the convective warming process. Here we address this zebrafish cryopreservation problem by using gold nanorods (GNRs) to assist in the warming process. Specifically, we microinjected the cryoprotectant propylene glycol into zebrafish embryos along with GNRs, and the samples were cooled at a rate of 90 000 °C/min in liquid nitrogen. We demonstrated the ability to unfreeze the zebrafish rapidly (1.4 × 107 °C/min) by irradiating the sample with a 1064 nm laser pulse for 1 ms due to the excitation of GNRs. This rapid warming process led to the outrunning of ice formation, which can damage the embryos. The results from 14 trials (n = 223) demonstrated viable embryos with consistent structure at 1 h (31%) and continuing development at 3 h (17%) and movement at 24 h (10%) postwarming. This compares starkly with 0% viability, structure, or movement at all time points in convectively warmed controls (n = 50, p < 0.001, ANOVA). Our nanoparticle-based warming process could be applied to the storage of fish, and with proper modification, can potentially be used for other vertebrate embryos.

The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acsnano.7b02216.

  • More information on the characterization of laser fluence rate and the selection of an appropriate GNR concentration; more details on thermal modeling setup that was used evaluating cooling and warming rates and thermal stress; data to support the success of laser GNR warming to rearm frozen HDF cells; histological images of zebrafish taken 7 days after being injected with GNRs (PDF)

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Received 30 March 2017
Date accepted 27 June 2017
Published online 13 July 2017
Published in print 22 August 2017
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