Research On High-speed Streaming Zebrafish Embryo Imaging Based On Linear CCD

The development of life sciences strongly depends on the miniature model organisms'development and utilization.However,the current used in miniature model organisms in the process of conducting scientific research mainly manual sorting screening and using traditional microscope imaging,this can't adapt to the large-scale genetic/drug screening requirement.The research topic of this paper is high-throughput imaging of zebrafish embryos using a linear?CCD-based flow imaging(Lc-FIS)system.The research can realize the function of high-throughput imaging of zebrafish embryos and real-time analysis of zebrafish embryos.The samples used in this study were zebrafish embryos,we designed two kinds of Iinear-CCD?based flow imaging system which aim at wild type zebrafish embryos and fluorescent zebrafish embryos,correspondingly.According to the demands,we built the linear?CCD-based flow imaging system.Lc-FIS consists of three modules:illumination module,microfluidic channel module and detecting module.The illumination module mainly includes laser,cylindrical mirror and other optical lens,a thin sheet of light was generated using a cylinder lens and illuminated on the samples to reduce the exposure of the living embryos to the laser.The liquid flow module mainly includes microfluidic channel,imaging capillary and other tube which is used to deliver zebrafish embryos.The imaging module is composed of tube lens,linear array CCD which is used to imaging for flowing zebrafish embryos with a high speed.Lc-FIS can image dozens of zebrafish embryos per second.An image algorithm was developed to recognize each embryo and to perform automatic analysis.For embryos with chorion,we use a Hough transformation algorithm to define a circle along the edge of the chorion.For embryos without chorion,we used edge detection and morphological operations to obtain the region of embryos from Lc-FIS images.We distinguished dead and living embryos according to the gray level distribution and conduced statistics of morphological characteristics of embryos at different growing stages.The research of this subject will actively promote the development of flow imaging system of large particle.Our linear-CCD-based flow imaging system(Lc-FIS)is promising for large-scale studies of the development of miniature model organisms in terms of various factors and drug discovery.