Integrated chip-based physiometer for automated fish embryo toxicity biotests in pharmaceutical screening and ecotoxicology

Akagi, J., Zhu, F., Hall, C.J., Crosier, K.E., Crosier, P.S., Wlodkowic, D.
Cytometry. Part A : the journal of the International Society for Analytical Cytology   85(6): 537-47 (Journal)
Registered Authors
Crosier, Phil, Hall, Chris
angiogenesis, drugs, fish embryo test, lab-on-a-chip, laboratory automation, microfluidics, transgenic models, zebrafish
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Drug Discovery
  • Ecotoxicology*/instrumentation
  • Ecotoxicology*/methods
  • Embryo, Nonmammalian/drug effects
  • Humans
  • Kinetics
  • Microfluidic Analytical Techniques/instrumentation
  • Microfluidic Analytical Techniques/methods
  • Pharmaceutical Preparations/administration & dosage*
  • Zebrafish*
24664821 Full text @ Cytometry A
Transgenic zebrafish (Danio rerio) models of human diseases have recently emerged as innovative experimental systems in drug discovery and molecular pathology. None of the currently available technologies, however, allow for automated immobilization and treatment of large numbers of spatially encoded transgenic embryos during real-time developmental analysis. This work describes the proof-of-concept design and validation of an integrated 3D microfluidic chip-based system fabricated directly in the poly(methyl methacrylate) transparent thermoplastic using infrared laser micromachining. At its core, the device utilizes an array of 3D micromechanical traps to actively capture and immobilize single embryos using a low-pressure suction. It also features built-in piezoelectric microdiaphragm pumps, embryo-trapping suction manifold, drug delivery manifold, and optically transparent indium tin oxide heating element to provide optimal temperature during embryo development. Furthermore, we present design of the proof-of-concept off-chip electronic interface equipped with robotic servo actuator driven stage, innovative servomotor-actuated pinch valves, and embedded miniaturized fluorescent USB microscope. Our results showed that the innovative device has 100% embryo-trapping efficiency while supporting normal embryo development for up to 72 hr in a confined microfluidic environment. We also showed data that this microfluidic system can be readily applied to kinetic analysis of a panel of investigational antiangiogenic agents in transgenic zebrafish lines. The optical transparency and embryo immobilization allow for convenient visualization of developing vasculature patterns in response to drug treatment without the need for specimen re-positioning. The integrated electronic interfaces bring the lab-on-a-chip systems a step closer to realization of complete analytical automation.
Genes / Markers
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes