Zebrafish embryo development in a microfluidic flow-through system

Eric M. Wielhouwer; Shaukat Ali; Abdulrahman Al-Afandi; Marko T. Blom; Marinus B. Olde Riekerink; Christian Poelma; Jerry Westerweel; Johannes Oonk; Elwin X. Vrouwe; Wilfred Buesink; Harald G. J. vanMil; Jonathan Chicken; Ronny van 't Oever; Michael K. Richardson

doi:10.1039/C0LC00443J

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Lab on a Chip

Issue 10, 2011

Miniaturisation for chemistry, physics, biology, materials science and bioengineering

Impact Factor 5.67 24 Issues per Year Indexed in MEDLINE

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Zebrafish embryo development in a microfluidic flow-through system

Eric M. Wielhouwer,^a Shaukat Ali,^a Abdulrahman Al-Afandi,^a Marko T. Blom,^b Marinus B. Olde Riekerink,^b Christian Poelma,^c Jerry Westerweel,^c Johannes Oonk,^b Elwin X. Vrouwe,^b Wilfred Buesink,^b Harald G. J. vanMil,^a Jonathan Chicken,^d Ronny van 't Oever^b and Michael K. Richardson*^a

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Institute of Biology, Leiden University, Sylvius Laboratory, Sylviusweg 72, 2333, BE, Leiden, The Netherlands
E-mail: richardsonmk@biology.leidenuniv.nl

Micronit Microfluidics BV, Enschede, The Netherlands

Laboratory for Aero & Hydrodynamics, Delft University of Technology, The Netherlands

FLIR Systems LTD, Nottingham, UK

Lab Chip, 2011,11, 1815-1824

DOI: 10.1039/C0LC00443J
Received 24 Sep 2010, Accepted 17 Mar 2011
First published online 14 Apr 2011

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The zebrafish embryo is a small, cheap, whole-animal model which may replace rodents in some areas of research. Unfortunately, zebrafish embryos are commonly cultured in microtitre plates using cell-culture protocols with static buffer replacement. Such protocols are highly invasive, consume large quantities of reagents and do not readily permit high-quality imaging. Zebrafish and rodent embryos have previously been cultured in static microfluidic drops, and zebrafish embryos have also been raised in a prototype polydimethylsiloxane setup in a Petri dish. Other than this, no animal embryo has ever been shown to undergo embryonic development in a microfluidic flow-through system. We have developed and prototyped a specialized lab-on-a-chip made from bonded layers of borosilicate glass. We find that zebrafish embryos can develop in the chip for 5 days, with continuous buffer flow at pressures of 0.005–0.04 MPa. Phenotypic effects were seen, but these were scored subjectively as ‘minor’. Survival rates of 100% could be reached with buffer flows of 2 µL per well per min. High-quality imaging was possible. An acute ethanol exposure test in the chip replicated the same assay performed in microtitre plates. More than 100 embryos could be cultured in an area, excluding infrastructure, smaller than a credit card. We discuss how biochip technology, coupled with zebrafish larvae, could allow biological research to be conducted in massive, parallel experiments, at high speed and low cost.

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