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Gold nanoparticle laser warming of cryopreserved zebrafish embryos

Award Information
Agency: Department of Health and Human Services
Branch: National Institutes of Health
Contract: 1R41OD024430-01
Agency Tracking Number: R41OD024430
Amount: $225,000.00
Phase: Phase I
Program: STTR
Solicitation Topic Code: OD
Solicitation Number: PA15-087
Solicitation Year: 2015
Award Year: 2017
Award Start Date (Proposal Award Date): 2017-05-01
Award End Date (Contract End Date): 2018-10-31
Small Business Information
San Diego, CA 92111-1806
United States
DUNS: 159070825
HUBZone Owned: No
Woman Owned: No
Socially and Economically Disadvantaged: No
Principal Investigator
 (858) 565-4227
Business Contact
Phone: (858) 565-4227
Research Institution
MINNEAPOLIS, MN 55455-0149
United States

 Nonprofit College or University

Project Summary Abstract
In the past decade laboratories around the world have produced tens of thousands of
mutant transgenic and wild type zebrafish lines for a wide range of genetic and biomedical
research Maintaining all of these valuable genotypes is expensive risky and beyond the
capacity of even the largest stock centers Cryopreservation of zebrafish sperm eggs and
embryos is vital to the strategy of NIH s Division of Comparative Medicine which envisions
increased multi institutional research using animal models To date zebrafish sperm and eggs
have been successfully cryopreserved but zebrafish embryos have not The main challenges
with zebrafish embryo cryopreservation are the large size of the embryo which limits the rate at
which the embryo can be externally cooled and warmed and a multi compartmental embryo
with different permeabilities preventing uniform diffusion of cryoprotectant agents Methods and
apparatus that enabled the long term storage and transport of cryopreserved embryos would
address a critical need for zebrafish researchers
This project will develop and optimize a gold nanoparticle GNP based rapid warming
technology that has successfully generated viable zebrafish grown from a cryopreserved
embryos Due to the large size of the embryo traditional warming mechanisms are too slow and
result in the formation of ice crystals that damage the embryos and prevent them from being
viable The key innovation of this project addresses this limitation in the warming step Injected
GNP act as a distributed network of ultra efficient heaters that generate warming rates of
millions of C min when illuminated with an infrared laser To achieve this ultrafast and
reproducible warming stable low toxicity GNPs with strong absorption at the laser wavelength
will be fabricated and their photothermal properties measured A micro injection technique
developed at the University of Minnesota will be optimized to circumvent the permeability barrier
to safely introduce cryoprotectant agents CPAs and GNPs into the yolk and chorion
compartments Embryos will be rapidly cooled to prevent damaging ice formation and laser
warming techniques will be optimized to increase embryo survival after thawing The combined
foundational research of Dr Bischof and his collaborators along with the expertise in GNP
synthesis and manufacturing at nanoComposix will allow for rapid optimization and
commercialization of this technology Project Narrative
Zebrafish are an important model organism with genes and organs that share many similarities
with humans allowing researchers to use zebrafish to study and better understand a cancer
and other diseases and test the efficacy of pharmaceuticals that could one day be used to treat
humans Researchers would benefit from a technology that allowed them to freeze and store
zebrafish embryos which has not been possible until now enabling improved storage
transport and sharing of zebrafish variants This collaborative effort between nanoComposix
and Prof John Bischof at the University of Minnesota seeks to optimize a new method enabling
rapid freezing and thawing of zebrafish embryos followed by expansion of the technology to
provide a universal platform for preservation of other organisms

* Information listed above is at the time of submission. *

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