Synthesis of lipid vesicles and other biological assays enabled by advanced rapid prototyping techniques

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Publication Type dissertation
School or College College of Engineering
Department Mechanical Engineering
Author Romanov, Valentin
Title Synthesis of lipid vesicles and other biological assays enabled by advanced rapid prototyping techniques
Date 2018
Description Liposomes are small vesicles filled with an aqueous solvent, bounded by a fluid shell that is comprised of a lipid bilayer. The bilayer may contain a single lipid species or a complex mixture that approximates the lipid content of a specific cellular membrane. This research developed a microfluidic and a biological framework for creating and probing the functionality of synthetic vesicles. To study vesicles that either approximated the size of cells or endosomal compartments, a microfluidic device was developed capable of creating nano- or microscale vesicles. This was the first demonstration of a microfluidic strategy able to access both size ranges. This technique enabled the discovery of novel functionality of several ESCRT-III (Endosomal Sorting Complex Required for Transport) proteins while also enabling the first in-depth electron microscopy analysis of liposomes created by a microfluidic platform. To encourage adoption of the microfluidic technique developed in this work, a three-dimensional (3D) printed framework was developed for manufacturing leak-free, transparent microfluidic devices. The methodology developed in this section led to the creation of the smallest ever FDM (Fused Deposition Modeling) 3D printed microfluidic channels (400 μm x 400 μm), demonstrating leak-free operation up to ~5MPa. To increase heat resistance of these devices, an annealing technique was developed for PLA (Poly(lactic Acid)). This was used to demonstrate the first DNA melting analysis on a 3D printed microfluidic device capable of withstanding temperatures up to 100ºC. iv A new method was developed for the 3D printing of 200 μm x 200 μm channels, the smallest to date using an FDM style printer. In conjunction with this method, a technique was developed for direct integration of a polycarbonate membrane into a 3D printed device. The entirely 3D printed protocol was used to demonstrate continuous formation of vesicles. This was the first demonstration of vesicles synthesized using a 3D printed platform. This technique enabled encapsulation of high ionic strength solutions in cell-sized vesicles (~5 μm), created in a low viscosity fluid.
Type Text
Publisher University of Utah
Dissertation Name Doctor of Philosophy
Language eng
Rights Management (c) Valentin Romanov
Format Medium application/pdf
ARK ark:/87278/s6sv3pk9
Setname ir_etd
Date Created 2021-06-16
Date Modified 2021-06-18
ID 1701773
Reference URL https://collections.lib.utah.edu/ark:/87278/s6sv3pk9
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