Batch Transfer of Zebrafish Embryos Into Multiwell Plates

Abstract

Abstract-This paper reports a prototype cooperative robotic system capable of transferring zebrafish embryos in parallel and depositing a single embryo per well in a 96-well microplate. A cell holding device was developed to trap multiple embryos in a regular pattern. The cell holding device and a microplate were positioned and aligned along multiple axes by the system. Embryo release strategies were systematically studied and compared. Experiments demonstrated that out of the 1056 zebrafish embryos used in experiments (i.e., 44 times parallel transfer into 11 96-well plates), 996 wells were successfully filled with one and only one zebrafish embryo, representing a success rate of 94.3%. Further experiments confirmed that the transferred embryos were able to develop into zebrafish with 100% survival rate.Note to Practitioners-Biological experiments and drug screen require the transfer of individual zebrafish embryos into standard multiwell microplates. Manually pipetting one and only one embryo into each well is tedious and time consuming. Different from the only commercial system for automated zebrafish embryo transfer, which is based on conventional, expensive flow cytometry, the proof-of-principle system reported in this paper utilizes a cell immobilization device and a cooperative robotic mechanism for parallel transfer of zebrafish embryos. The system architecture and transfer techniques promise efficient, cost-effective zebrafish embryo transfer for applications requiring molecule tests on a high number of cells.Index Terms-Batch transfer, control and alignment, cooperative robotic mechanism, drug screen, high-throughput screen (HTS), multiwell microplate, zebrafish embryo.

Evaluation of the Effect of AqueousMomordica charantiaLinn. Extract on Zebrafish Embryo Model through Acute Toxicity Assay Assessment

“…One zebrafish embryo was transferred into each of the 96-well plates using a pasteur pipette at 24 hours post-fertilization (24hpf) [21]. Next, the treatment was given by pipetting the sample into each well in the 96-well plates based on different series of concentrations.…”

Embryotoxicity and Teratogenic Effects of Parmotrema Tinctorum (Nyl) Hale on Zebrafish (Danio Rerio) and Cytotoxic Activity of Lichen Extract on Hek293t Cell Line

“…The Zebrafishes were fed with commercial feed five times per day. After selection, at 24-hour postfertilization, using a Pasteur pipette the embryos were transferred into each of the 96 well plates [20].…”