Development of a nanotechnology system for the dynamic quantification of cell-produced single molecules with spatial resolution during homeostasis and stress

Cellular behaviour during homeostasis and disease is steered by a balanced interaction between a set of cell populations and their secreted signalling molecules. To understand these processes, dynamic monitoring of the production of specific factors such as growth factors or inflammatory cytokines can help to understand when a cellular system is entering a compromised stage, such as in disease. The ability to detect these factors at the micro tissue or single cell level would not only help to understand what happens during the early stage of the disease, but also improve our ability to detect a failing system at an earlier stage. 

This Master Thesis project aims to develop a quantitative system to enable the dynamic real-time measurement of molecules with spatial resolution related to the producing cell, during homeostasis and upon induced stress. This will be done by implementing a membrane structure to guide cell-secreted molecules into micro-compartments where the dynamic quantification of the specific molecules will be quantified using biological nanopores. As a steppingstone, during this thesis we will aim to have the cells and microcompartments integrated, while looking at cell secretion using fluorescent techniques. More specifically, during this thesis project, the student will start to optimize the fluorescent assays to detect specific molecules, quantify molecule transfer over dedicated nanopore membranes, and explore options to integrate cells, membranes and micro-wells towards a first proof-of-concept system.