Automated Lab-on-a-Chip Technology for Fish Embryo Toxicity Tests Performed under Continuous Microperfusion (μFET).
The fish embryo toxicity (FET) biotest has gained popularity as one of the alternative approaches to acute fish toxicity tests in chemical hazard and risk assessment. Despite the importance and common acceptance of FET, it is still performed in multiwell plates and requires laborious and time-consuming manual manipulation of specimens and solutions. This work describes the design and validation of a microfluidic Lab-on-a-Chip technology for automation of the zebrafish embryo toxicity test common in aquatic ecotoxicology. The innovative device supports rapid loading and immobilization of large numbers of zebrafish embryos suspended in a continuous microfluidic perfusion as a means of toxicant delivery. Furthermore, we also present development of a customized mechatronic automation interface that includes a high-resolution USB microscope, LED cold light illumination, and miniaturized 3D printed pumping manifolds that were integrated to enable time-resolved in situ analysis of developing fish embryos. To investigate the applicability of the microfluidic FET (μFET) in toxicity testing, copper sulfate, phenol, ethanol, caffeine, nicotine, and dimethyl sulfoxide were tested as model chemical stressors. Results obtained on a chip-based system were compared with static protocols performed in microtiter plates. This work provides evidence that FET analysis performed under microperfusion opens a brand new alternative for inexpensive automation in aquatic ecotoxicology.
Feng
F
Zhu
Université de Lyon, UMR LEHNA 5023, USC INRA, ENTPE, rue Maurice Audin, Vaulx-en-Velin F-69518, France.
Adriana
A
Wigh
ARMI, Monash University , Wellington Road, Clayton, VIC 3800, Australia.
Timo
T
Friedrich
Université de Lyon, UMR LEHNA 5023, USC INRA, ENTPE, rue Maurice Audin, Vaulx-en-Velin F-69518, France.
Alain
A
Devaux
Université de Lyon, UMR LEHNA 5023, USC INRA, ENTPE, rue Maurice Audin, Vaulx-en-Velin F-69518, France.
Sylvie
S
Bony
Ecotoxicology Research Group, School of Applied Sciences, RMIT University , Bowen Street, Melbourne, VIC 3001, Australia.
Dayanthi
D
Nugegoda
ARMI, Monash University , Wellington Road, Clayton, VIC 3800, Australia.
Jan
J
Kaslin
Centre for Additive Manufacturing, RMIT University , 58 Cardigan Street, Melbourne, VIC 3053, Australia.
Donald
D
Wlodkowic
D000818:Animals; D002110:Caffeine / Q000633:toxicity; D019327:Copper Sulfate / Q000633:toxicity; D004121:Dimethyl Sulfoxide / Q000633:toxicity; D004625:Embryo, Nonmammalian / Q000187:drug effects; D000431:Ethanol / Q000633:toxicity; D056656:Lab-On-A-Chip Devices; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D009538:Nicotine / Q000633:toxicity; D066330:Printing, Three-Dimensional; D018675:Toxicity Tests / Q000295:instrumentation* / Q000379:methods; D015027:Zebrafish / Q000196:embryology*
D002110:Caffeine; D000431:Ethanol; D009538:Nicotine; D019327:Copper Sulfate; D004121:Dimethyl Sulfoxide