TitleAutomated zebrafish chorion removal and single embryo placement: optimizing throughput of zebrafish developmental toxicity screens.
Publication TypeJournal Article
Year of Publication2012
AuthorsMandrell, D, Truong, L, Jephson, C, Sarker, MR, Moore, A, Lang, C, Simonich, SLMassey, Tanguay, RL
JournalJ Lab Autom
Volume17
Issue1
Pagination66-74
Date Published2012 Feb
ISSN2211-0690
KeywordsAnimals, Automation, Laboratory, Chorion, Drug Discovery, Drug Evaluation, Preclinical, Embryonic Development, High-Throughput Screening Assays, Robotics, Single Embryo Transfer, Toxicity Tests, Zebrafish
Abstract

The potential of the developing zebrafish model for toxicology and drug discovery is limited by inefficient approaches to manipulating and chemically exposing zebrafish embryos-namely, manual placement of embryos into 96- or 384-well plates and exposure of embryos while still in the chorion, a barrier of poorly characterized permeability enclosing the developing embryo. We report the automated dechorionation of 1600 embryos at once at 4 h postfertilization (hpf) and placement of the dechorionated embryos into 96-well plates for exposure by 6 hpf. The process removed ≥95% of the embryos from their chorions with 2% embryo mortality by 24 hpf, and 2% of the embryos malformed at 120 hpf. The robotic embryo placement allocated 6-hpf embryos to 94.7% ± 4.2% of the wells in multiple 96-well trials. The rate of embryo mortality was 2.8% (43 of 1536) from robotic handling, the rate of missed wells was 1.2% (18 of 1536), and the frequency of multipicks was <0.1%. Embryo malformations observed at 24 hpf occurred nearly twice as frequently from robotic handling (16 of 864; 1.9%) as from manual pipetting (9 of 864; 1%). There was no statistical difference between the success of performing the embryo placement robotically or manually.

DOI10.1177/2211068211432197
Alternate JournalJ Lab Autom
PubMed ID22357610
PubMed Central IDPMC3327291
Grant ListP30 ES000210-37 / ES / NIEHS NIH HHS / United States
R01 ES016896 / ES / NIEHS NIH HHS / United States
F31 ES019445-02 / ES / NIEHS NIH HHS / United States
RC4 ES019764-01 / ES / NIEHS NIH HHS / United States
F31 ES019445 / ES / NIEHS NIH HHS / United States
RC4 ES019764 / ES / NIEHS NIH HHS / United States
P42 ES016465 / ES / NIEHS NIH HHS / United States
R01 ES016896-04 / ES / NIEHS NIH HHS / United States
P30 ES000210 / ES / NIEHS NIH HHS / United States