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

Journal of Laboratory Automation
David MandrellRobert L Tanguay

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.

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Citations

Jun 14, 2013·Zebrafish·Ralf MikutNadine Peyriéras
Apr 4, 2013·Toxicological Sciences : an Official Journal of the Society of Toxicology·Ki-Tae KimRobert L Tanguay
Oct 10, 2012·Future Medicinal Chemistry·Jonathan R MathiasJeff S Mumm
Jan 17, 2014·Diseases of Aquatic Organisms·Michael L KentRobert L Tanguay
May 21, 2013·Toxicology and Applied Pharmacology·Andrea L KnechtRobert L Tanguay
Feb 14, 2015·Current Environmental Health Reports·Sean M BugelAntonio Planchart
Jun 2, 2015·Toxicology Reports·Leah C WehmasRobert L Tanguay
Aug 28, 2015·Journal of Pharmacological and Toxicological Methods·Steven CassarTodd Cole
Feb 12, 2014·Alcoholism, Clinical and Experimental Research·George R FlentkeSusan M Smith
Oct 6, 2015·Journal of Nanoparticle Research : an Interdisciplinary Forum for Nanoscale Science and Technology·Charles GeraciSara Brenner
Jul 5, 2015·Journal of Environmental Sciences (China)·M Martínez-SalesF J Espinós
Aug 2, 2015·Biochimica Et Biophysica Acta·John J StegemanPhilippe Urban
Feb 18, 2014·Anatomia, histologia, embryologia·Y ComabellaT García-Galano
Dec 3, 2014·Current Opinion in Chemical Biology·Andrew J Rennekamp, Randall T Peterson
Dec 19, 2014·Environmental Health and Toxicology·Ki-Tae Kim, Robert L Tanguay
Jun 13, 2015·Journal of Nanoparticle Research : an Interdisciplinary Forum for Nanoscale Science and Technology·Bryan HarperStacey Harper
Apr 17, 2015·Toxicological Sciences : an Official Journal of the Society of Toxicology·Julieta M Panzica-KellyKaren A Augustine-Rauch
May 10, 2013·Toxicology and Applied Pharmacology·Britton C GoodaleRobert L Tanguay
Mar 8, 2017·The Journal of Nutritional Biochemistry·Laura M BeaverEmily Ho
May 10, 2017·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Guzmán ÁlvarezMercedes González
Dec 24, 2017·Environmental Toxicology and Chemistry·Lauren A KristofcoBryan W Brooks
Jan 17, 2014·Zebrafish·Sridhara ChakravarthySunil Kumar Sukumaran
Apr 25, 2017·Nature Nanotechnology·Santiswarup SinghaPere Santamaria
Jun 19, 2013·Green Chemistry : an International Journal and Green Chemistry Resource : GC·Ki-Tae Kim, Robert L Tanguay
Feb 12, 2017·Toxicological Sciences : an Official Journal of the Society of Toxicology·Anna C ChlebowskiRobert L Tanguay
Oct 19, 2013·Toxicological Sciences : an Official Journal of the Society of Toxicology·Lisa TruongRobert L Tanguay

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Methods Mentioned

BETA
dissecting
transgenic
gene knockdown

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