This thesis presents the design, development and application of several platforms through which to generate giant vesicles for biophysical and mechanical membrane studies. There has been a growing focus on manufacturing model membrane systems with improved throughput and structural properties in recent years. GUVs are a popular model membrane system for studying lipid membrane-associated phenomena due to their inherent similarity to biological cells. Traditional methods to construct vesicles offer little control over nuanced membrane properties such as asymmetry and patterning, which has paved the way for more refined techniques to be developed.
This thesis details the development of a microfluidic platform technology that addresses this chasm in sophisticated GUV fabrication strategies. The technique presented offers control over key structural features such as vesicle size dispersity, internal content, membrane composition and asymmetry. Vesicles were investigated using contour detection and fluctuation analysis in order to quantify the bending rigidity in membranes constructed by microfluidics for the very first time.
Furthermore, the emulsion phase transfer (EPT) method was refined for the construction of GUVs with phase separated membranes across three of compositions. This is the first investigation concerning domain formation in membranes constructed from emulsion precursors at a range of a compositions.
The progress made in advancing platform technologies opens up various avenues through which to further explore biophysical phenomena such as a lipid flip-flop dynamics, as well as for the high-throughput generation of artificial cell systems, with potential relevance for therapeutic applications such as smart drug delivery.