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Title: Single cell analysis and cell sorting using microfluidic devices with application to circulating tumour cells
Author: Kaplinsky, Joseph John
ISNI:       0000 0004 2714 7934
Awarding Body: Imperial College London
Current Institution: Imperial College London
Date of Award: 2012
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This thesis describes the development of integrated microfluidic technology for single cell proteomic analysis, focusing on circulating tumour cells (CTCs). While single cell proteomic analysis has wide applicability across biology and medicine, CTCs form an ideal first application. Circulating tumour cells are intimately involved in metastasis, the step in cancer overwhelmingly responsible for death, yet have proved hard to study. Single cell microfluidic technology is ideal first because the quantity of material available is inherently at the level of a few cells and second because cell to cell variation is of great interest. Chapter 1 is an introduction to the field. In chapter 2 a microfluidic sandwich assay for quantification of protein at the single cell level is described. In chapter 3 the isolation of CTCs in a microfluidic device is described. This relies on taking the output of the CellSearch® system and inputing it to a microfluidic device. While CTCs were identified, the result showed that a more systematic approach is required for counting and integration with the single cell assay previously described. Chapters 4 and 5 describe development of technology suitable for counting and isolation of CTCs integrated into a microfluidic device with single cell proteomic analysis, although the work done here makes use of fluorescently labelled beads and model cell lines rather than CTCs from patient samples. Chapter 4 describes microfluidic cytometry that can be used to count and identify a labelled population of cells, such as stained CTCs. Chapter 5 describes the prelimary development of a sorting system suitable for isolation of CTCs integrated with the cytometer.
Supervisor: de Mello, Andrew ; Ces, Oscar ; Klug, David Sponsor: EPSRC
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral