Image-Based Fluidic Sorting System for Automated Zebrafish Egg Sorting into Multiwell Plates

Abstract

T he global demand for the reduction of animal testing has led to the emergence of Zebrafish eggs/larvae as model organisms to replace current adult animal testing in, for example, toxicity testing. Because of the egg size (diameter 1.6 mm) and the relatively easy maintenance of Zebrafish farms the eggs also offer high-throughput screening (HTS). However, the current bottleneck for HTS is the cost-efficient placing of individual organisms into single wells of a multiwell plate (MWP). The system presented here is capable of storing, sorting, and placing individual organisms in a highly reproducible manner. In about 11 min a complete 96-MWP is filled, which corresponds to about 8 sec per egg. The survival rate of fertilized transgenic and wild-type eggs was comparable to the one of the control (control 6.7%, system 7.6%). Furthermore, it was also possible to place dechorionated eggs into individual wells. The results demonstrate that the cost efficient system works gentle and reliable enough to disburden scientists from the exhausting and monotonous job of placing single eggs into single wells, such that they can concentrate on the scientific aspects of their experiments and create results with a higher statistical relevance.

A Chemical Screening Procedure for Glucocorticoid Signaling with a Zebrafish Larva Luciferase Reporter System

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Automated Processing of Zebrafish Imaging Data: A Survey

“…Relatively simple real-time image processing routines are applied to support, for example, automated sorting or selective image acquisition. Examples include the development of flowbased systems for high-throughput imaging and laser microsurgery of larvae ''on the fly,'' 59 workflows for the automatic injection of bacteria into the yolk, 22 image-based sorting robots, 33,61 a screening platform with automated manipulation of zebrafish including orienting and positioning regions of interest within the microscope's field of view, 25 and automated zooming into ROI for heartbeat detection. 67 …”

The developing zebrafish (Danio rerio): A vertebrate model for high‐throughput screening of chemical libraries

“…Devices that automate the embryo or larvae handling and placement into wells are available commercially (CellBot, Centre Suisse d'Electronique et de Microtechnique; COPAS and BioSorter, Union Biometrica, Holliston, MA), and recently a small tabletop design has been reported (Graf et al, 2011). Robotic pipettors are also commercially available to provide the correct volume of media per well and to apply drug doses either as small liquid additions or from array pins that transfer drugs from resource dosing plates.…”