Micromanipulation System for Isolating a Single Cryptosporidium Oocyst

In this paper, an integrated system for contact micromanipulation of Cryptosporidium oocysts is presented. The system integrates five actuators and a partially automated control system and contacts the oocyst using a drawn glass end effector with tip dimensions of 1 μ m. The system is intended to allow single cell analysis (SCA) of Cryptosporidium-a very harmful parasite found in water supplies-by isolating the parasite oocyst of 5 μ m diameter in a new environment. By allowing this form of analysis, the source of Cryptosporidium can be found and potential harm to humans can be reduced. The system must overcome the challenges of locating the oocysts and end effector in 3D space and contact adhesion forces between them, which are prominent over inertial forces on this scale. An automated alignment method is presented, using the Prewitt operator to give feedback on the level of focus and this system is tested, demonstrating alignment accuracy of <2 μ m. Moreover, to overcome the challenge of adhesion forces, use of dry and liquid environments are investigated and a strategy is developed to capture the oocyst in the dry environment and release in the liquid environment. An experiment is conducted on the reliability of the system for isolating a Cryptosporidium oocyst from its culture, demonstrating a success rate of 98%.