A microfluidic on-chip imaging cell sorter has several advantages over conventional cell sorting methods, especially to identify cells with complex morphologies such as clusters. One of the remaining problems is how to efficiently discriminate targets at the species level without labelling. Hence, we developed a label-free microfluidic droplet-sorting system based on image recognition of cells in droplets. To test the applicability of this method, a mixture of two plankton species with different morphologies (Dunaliella tertiolecta and Phaeodactylum tricornutum) were successfully identified and discriminated at a rate of 10 Hz. We also examined the ability to detect the number of objects encapsulated in a droplet. Single cell droplets sorted into collection channels showed 91 ± 4.5% and 90 ± 3.8% accuracy for D. tertiolecta and P. tricornutum, respectively. Because we used image recognition to confirm single cell droplets, we achieved highly accurate single cell sorting. The results indicate that the integrated method of droplet imaging cell sorting can provide a complementary sorting approach capable of isolating single target cells from a mixture of cells with high accuracy without any staining.

The purpose of this study was the development of a digital water color measurement system using a CCD optical device. Photographs of the standard medium of Forel and Ule water color scales in the laboratory, and one of sea surface above a Secchi disc (Z=SD/2) immersed in seawater were taken. The colors of these pictures were estimated with the value of CIE L*a*b*. Water color assessment was conducted with a digital photo-image. In the case of the Forel scale (No. 1-11), the b* value was so high that the water color number was large. In the Ule scale , the a* value became high, which is why the number on the water color scale was large. The color of these pictures showed that the a* value in the Forel scale and the b* value in the Ule scale increase with the increase of the F value. The a* value of seawater color was always lower than the one on the Forel and Ule water color scales. This indicates that the color of the scales differs from actual seawater color. It was concluded that water color number can more effectively be assessed by estimating the ΔEab* color difference between the water scales and actual seawater color.