Development of an automated somatic embryo production system equipped with image analysis facilities

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The objective of the study was to develop an automated somatic embryo production system, in which good quality torpedo stage embryos were selected for further treatment of transplant or artificial seed production. The selection was executed based on image analysis of each somatic embryo. The study consisted of three parts.1. Development of an automated embryogenic callus selection system. In a conventional manual method of somatic embryo production, embryogenic callus was selected by sieving with three different size mesh. This procedure was automated and tested using carrot callus. The results showed increased ratio of malformed embryos by mechanical sieving, but that the automation was possible.2. Development of image analysis method of somatic embryos. Image analysis algorithms for selecting quality torpedo stage embryos were developed and tested. Algorithms based on thinning, generalized Hough transform, and Fourier descriptor were compared using carrot somatic embryo images. The results showed that the thinning algorithm was most promising which gave results 90% in agreement with human judgments.3. Development of SOMES (SOMatic Embryo Sorter). A system for selecting embryos was constructed and tested. The thinning algorithm was implemented. Path of liquid medium containing embryos was changed according to the results of image analysis of each embryo. The results showed the effectiveness of SOMES, but at the same time difficulties in image analysis of moving embryos in the imaging cell. The reasons of misjudgments were analyzed and points to be improved were suggested : use of a thinner tube for the imaging cell, and acquiring images from two angles. Efforts are being made to improve SOMES in this direction.