Axial orientation control of zebrafish larvae using artificial cilia

  • Chia-Yuan ChenEmail author
  • Tsung-Chun Chang Chien
  • Karthick Mani
  • Hsiang-Yu Tsai
Research Paper


Zebrafish has been used as an important vertebrate model of genetic screening and new drug development because of excellent characteristics, such as optical transparency, rapid ex vivo growth, and high genetic similarity to humans. Despite these advantages, studies on zebrafish are limited because of the lack of a robust and reliable method to manipulate zebrafish during microinjection and screening, as well as time-lapse imaging. In this work, a new microfluidic concept that utilizes a series of magnetically actuated artificial cilia integrated into a microchannel was employed to control the orientation of zebrafish larvae with a validated axial rotation capability. In contrast to conventional methods, the proposed method enables a highly accurate small-angle (0°–20°) stepwise axial rotation of a larva inside the microchannel with less detrimental effects on larval growth. The hemodynamics in a selected vessel was then imaged during the axial rotation of the tested larva to assist cardiovascular assessment. In addition, the bioactivity of the tested larvae remains stable without short-term negative effects after the imaging. The proposed platform, along with the provided analytical paradigm, can facilitate future zebrafish screening via microfluidics in the pharmaceutical industry.


Orientation control Zebrafish Artificial cilia Microfluidics 



This study was supported by Ministry of Science and Technology of Taiwan under Contracts Nos. MOST 104-2221-E-006-169 and MOST 102-2221-E-006-297-MY3 (to Chia-Yuan Chen). This work would not be possible without the facility provided by Center for Micro/Nano Science and Technology, National Cheng Kung University. The research was in part supported by the Ministry of Education, Taiwan, R.O.C., through the Aim for the Top University Project to National Cheng Kung University (NCKU).

Supplementary material

10404_2015_1668_MOESM1_ESM.docx (138 kb)
Supplementary material 1 (DOCX 138 kb)

Supplementary material 2 (MP4 9288 kb)


  1. Bischel LL, Mader BR, Green JM, Huttenlocher A, Beebe DJ (2013) Zebrafish Entrapment By Restriction Array (ZEBRA) device: a low-cost, agarose-free zebrafish mounting technique for automated imaging. Lab Chip 13:1732–1736CrossRefGoogle Scholar
  2. Brown D (2013) Sharing video experiments with tracker digital libraries paper presented at the American Association of Physics Teachers, AAPT, New orleans, LouislanaGoogle Scholar
  3. Chang TY, Pardo-Martin C, Allalou A, Wahlby C, Yanik MF (2012) Fully automated cellular-resolution vertebrate screening platform with parallel animal processing. Lab Chip 12:711–716CrossRefGoogle Scholar
  4. Chen CY, Cheng CM (2014) Microfluidics expands the zebrafish potentials in pharmaceutically relevant screening. Adv Healthc Mater 3:940–945CrossRefGoogle Scholar
  5. Chen CY, Patrick MJ, Corti P, Kowalski W, Roman BL, Pekkan K (2011) Analysis of early embryonic great-vessel microcirculation in zebrafish using high-speed confocal muPIV. Biorheology 48:305–321Google Scholar
  6. Chen CY, Anton R, Hung MY, Menon P, Finol EA, Pekkan K (2014) Effects of intraluminal thrombus on patient-specific abdominal aortic aneurysm hemodynamics via stereoscopic particle image velocity and computational fluid dynamics modeling. J Biomech Eng 136:031001CrossRefGoogle Scholar
  7. Choudhury D et al (2012) Fish and chips: a microfluidic perfusion platform for monitoring zebrafish development. Lab Chip 12:892–900CrossRefGoogle Scholar
  8. Funfak A, Brosing A, Brand M, Kohler JM (2007) Micro fluid segment technique for screening and development studies on Danio rerio embryos. Lab Chip 7:1132–1138CrossRefGoogle Scholar
  9. Hedrick TL (2008) Software techniques for two- and three-dimensional kinematic measurements of biological and biomimetic systems. Bioinspir Biomim 3:034001CrossRefGoogle Scholar
  10. Kaufmann A, Mickoleit M, Weber M, Huisken J (2012) Multilayer mounting enables long-term imaging of zebrafish development in a light sheet microscope. Development 139:3242–3247CrossRefGoogle Scholar
  11. Lin X et al (2014) High-throughput mapping of brain-wide activity in awake and drug-responsive vertebrates. Lab Chip 15:680–689CrossRefGoogle Scholar
  12. Malone MH, Sciaky N, Stalheim L, Hahn KM, Linney E, Johnson GL (2007) Laser-scanning velocimetry: a confocal microscopy method for quantitative measurement of cardiovascular performance in zebrafish embryos and larvae. BMC Biotechnol 7:40CrossRefGoogle Scholar
  13. Pardo-Martin C, Chang TY, Koo BK, Gilleland CL, Wasserman SC, Yanik MF (2010) High-throughput in vivo vertebrate screening. Nat Methods 7:634–636CrossRefGoogle Scholar
  14. Tamplin OJ, Zon LI (2010) Fishing at the cellular level. Nat Methods 7:600–601CrossRefGoogle Scholar
  15. Wielhouwer EM et al (2011) Zebrafish embryo development in a microfluidic flow-through system. Lab Chip 11:1815–1824CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Chia-Yuan Chen
    • 1
    Email author
  • Tsung-Chun Chang Chien
    • 1
  • Karthick Mani
    • 1
  • Hsiang-Yu Tsai
    • 1
  1. 1.Department of Mechanical EngineeringNational Cheng Kung UniversityTainanTaiwan

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