Isolation of Parasites by Micropipette Aspiration

This study presents the preliminary development of separating parasites with a robotics approach. Cryptosporidium is a parasite that causes sickness in people and animals. Cryptosporidium oocysts, which are approximately 5 mu m in size, contain four haploid sporozoites. It is currently unproven whether these 'siblings' are genetically identical or not. Common methods of selective cell isolation were considered for separation and the pick-and-place with microaspiration technique was selected due to the ability to work with low volume samples that would be the four sporozoites from a single oocyst. The drawbacks of high labour and time cost were offset by considering automation. The forming of the micropipettes was developed to produce the best tip for separating the oocysts and sporozoites. The optimum flow rate of the syringe pumps and the optimum angle of the micropipettes during microaspiration were intensively tested and found to be 12 mu l/min and 45 degrees respectively. The manual process was able to separate the Cryptosporidium oocyst and sporozoites. The separation process was found to be possible to automate with separating silica microbeads using computer vision. Overall, the findings of this paper are that parasites can be separated with micro-aspiration approach and the process can be automated to make the process viable within laboratories.