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
