OpenSource Lab‐on‐a‐Chip Physiometer for Accelerated Zebrafish Embryo Biotests

Abstract

Zebrafish (Danio rerio) embryo assays have recently come into the spotlight as convenient experimental models in both biomedicine and ecotoxicology. As a small aquatic model organism, zebrafish embryo assays allow for rapid physiological, embryo‐, and genotoxic tests of drugs and environmental toxins that can be simply dissolved in water. This protocol describes prototyping and application of an innovative, miniaturized, and polymeric chip‐based device capable of immobilizing a large number of living fish embryos for real‐time and/or time‐lapse microscopic examination. The device provides a physical address designation to each embryo during analysis, continuous perfusion of medium, and post‐analysis specimen recovery. Miniaturized embryo array is a new concept of immobilization and real‐time drug perfusion of multiple individual and developing zebrafish embryos inside the mesofluidic device. The OpenSource device presented in this protocol is particularly suitable to perform accelerated fish embryo biotests in ecotoxicology and phenotype‐based pharmaceutical screening. Curr. Protoc. Cytom. 67:9.44.1‐9.44.16. © 2014 by John Wiley & Sons, Inc.

Genotoxicity of Nanomaterials: Advanced In Vitro Models and High Throughput Methods for Human Hazard Assessment—A Review

“…The literature search resulted in 59 papers for “genotoxicity & 3D in vitro model”, 73 papers for “genotoxicity & advanced in vitro model”, 134 papers for “genotoxicity & high throughput”, eight papers for “genotoxicity & high throughput & nanomaterials” [ 15 , 16 , 17 , 18 , 19 , 20 , 21 ] and six papers for “genotoxicity & high throughput & nanoparticles” (4 are the same as for “genotoxicity & high throughput & nanomaterials”) [ 16 , 17 , 18 , 19 , 21 , 22 ], seven papers for “genotoxicity & organ on chip“ [ 23 , 24 , 25 , 26 , 27 , 28 , 29 ], seven papers for “genotoxicity & 3D models & nanoparticles” [ 30 , 31 , 32 , 33 , 34 , 35 , 36 ], 11 papers for “genotoxicity & 3D models & nanomaterials” [ 30 , 31 , 33 , 36 , 37 , 38 , 39 , 40 , 41 ] for the period of 20 years (2000–2020), whereby the year 2020 was considered only from January to August. ( Table 1 ).…”

“…If genotoxicity is measured after, e.g., three and 24 h exposure, as is common for the comet assay, then cytotoxicity should also be measured after three and 24 h’ exposure. The alamarBlue™ assay is a convenient and reliable cytotoxicity assay to be used in combination with the comet assay, which has been tested and found to be applicable with several different NMs [ 6 , 26 , 36 ].…”

Microfluidics: reframing biological enquiry

“…Interestingly, the WUSCHEL-RELATED HOMEOBOX 5 (WOX5) promoter undergoes a burst of activity in the elongation zone of the seedling, producing a standing wave of WOX5 expression in that region, which indicates that there is a self-sustaining regulatory network for WOX5 expression 67 . Similarly, microfluidic systems are being developed to culture arrays of zebrafish embryos 70 as a high-throughput in vivo platform for toxicology and drug screening 69,71 .…”

Microfluidic device for a rapid immobilization of Zebrafish larvae in environmental scanning electron microscopy

“…This popularity stems from the fact that both zebrafish and Xenopus provide some unique investigative advantages as compared with bioassays that employ cell lines, isolated primary cells, and/or tissue samples . The main advantage is the potential for miniaturisation and automation of experiments, while still allowing for effects under study to occur in the context of intact physiological milieu, with all its intercellular, multiorgan, and multisystem interactions . Therefore, they bridge the existing gap between the traditional high‐throughput cell‐based in vitro assays and low‐throughput rodent in vivo tests.…”

“…To facilitate high‐throughput studies using small model organisms, a new generation of miniaturized lab‐on‐a‐chip (LOC) devices have recently been developed for both zebrafish and Xenopus embryo on‐chip culture and pharmacological interrogation . Only recently, Akagi et al reported on applications of microfluidic living embryo arrays for hydrodynamic trapping of single living zebrafish embryos and larvae . These specimens are intrinsically challenging to image due to immobilisation difficulties and rapid dislodgement related to high momentums of inertia.…”