PUBLICATION

Automated zebrafish chorion removal and single embryo placement: optimizing throughput of zebrafish developmental toxicity screens

Authors
Mandrell, D., Truong, L., Jephson, C., Sarker, M.R., Moore, A., Lang, C., Simonich, M.T., and Tanguay, R.L.
ID
ZDB-PUB-120227-6
Date
2012
Source
Journal of Laboratory Automation   17(1): 66-74 (Journal)
Registered Authors
Tanguay, Robyn L.
Keywords
zebrafish, automation, chorion, robotic
MeSH Terms
  • Animals
  • Automation, Laboratory*
  • Chorion/metabolism*
  • Drug Discovery*
  • Drug Evaluation, Preclinical/instrumentation
  • Drug Evaluation, Preclinical/methods
  • Embryonic Development/drug effects
  • High-Throughput Screening Assays*
  • Robotics/trends
  • Single Embryo Transfer/instrumentation*
  • Single Embryo Transfer/methods
  • Toxicity Tests*
  • Zebrafish
PubMed
22357610 Full text @ J. Lab. Autom.
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.

Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping