Density-Based Separations in Aqueous Multiphase Systems: Tools for Biological Research and Low-Cost Diagnostics
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CitationKumar, Ashok Ashwin. 2014. Density-Based Separations in Aqueous Multiphase Systems: Tools for Biological Research and Low-Cost Diagnostics. Doctoral dissertation, Harvard University.
AbstractCells often exist in heterogeneous mixtures. Density provides a property to separate several types of cells from the mixed sample in which they originate. Density-based separation methods provide a standard method to quickly separate or enrich specific populations of cells, such as lymphocytes from whole blood. This dissertation explores the use of aqueous multiphase systems (AMPS) as self-forming step-gradients in density for the separation of cells. AMPS were first discovered over a hundred years ago as aqueous two-phase systems. Density as a tool to separate cells is at least as old. Despite this long history, the work in this thesis is the first work to use AMPS to perform density-based separations on cells. This combination provides a powerful technique to separate cells and enable new testing at the point-of-care. Chapter 1 provides a short overview of aqueous multiphase systems and density-based separations of cells. Chapter 2 describes the process of taking technology, including AMPS, from a demonstration in a laboratory to a large scale evaluation in a field setting. In Chapter 3 and Appendix I, AMPS provide a means to enrich reticulocytes from whole blood as a means to grow malaria parasites. Chapter 4 and Appendix II describe the development and proof-of-prinicple of a density-based diagnostic test for sickle cell disease (SCD) using AMPS. Chapter 5 and Appendix III detail the results of a large scale field evaluation of a rapid test for SCD using AMPS in Zambia. Demonstrations of AMPS for density- and size-based separations are provided in Appendices IV and V. Appendix VI demonstrates the general usefulness of density to separate crystal polymorphs with another density-based separation method: magnetic levitation in a paramagnetic fluid. Beyond density, novel combinations of technology, such as electrochemistry and telecommunications provide opportunities for enabling global health (Appendix VII).
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