An integrated system for selection and manipulation of single cells in culture
Author(s)Guggenheim, Jacob (Jacob William)
Massachusetts Institute of Technology. Department of Mechanical Engineering.
H. Harry Asada.
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Cellular heterogeneity plays a critical role in processes like embryonic development, drug-resistance, and immune response. The ability to select and manipulate individual cells greatly increases the ability to study heterogeneity within cultures. As such, a number of different single cell sorting techniques have been developed. While filling their respective niches, there does not currently exist a system capable of sorting individual cells based upon an arbitrary set of morphological or otherwise observable features while providing good enough throughput to deal with working within biologically relevant timescales. To address this need, an integrated system for the selection and manipulation of single cells in 2D culture was developed by automating both the selection of cells using computer vision and manipulation of cells using micro-capillaries. This system was then applied to study intestinal organoid formation. Specifically, current protocols for the production of intestinal organoids suffer from low yield, with only ~12% of the highly heterogeneous nascent spheroids successfully maturing into intestinal organoids. It was found that by sorting for nascent spheroids that had an effective diameter greater than 75[mu]m and had an inner mass, the maturation percentage was increased to ~40%. Separately, the system was fitted with a microfluidic device capable of producing a local trypsin zone within 2D culture and single adherent cells were lifted. This shows promise for expanding the set of applications that single cell sorting can be applied.
Thesis: S.M., Massachusetts Institute of Technology, Department of Mechanical Engineering, 2016.Cataloged from PDF version of thesis.Includes bibliographical references (pages 35-36).
DepartmentMassachusetts Institute of Technology. Department of Mechanical Engineering.
Massachusetts Institute of Technology