A microfluidic approach for biological applications using vertical 3D nanostructures

TY - BOOK

T1 - A microfluidic approach for biological applications using vertical 3D nanostructures

AU - Aliti, Liridon

PY - 2018

Y1 - 2018

N2 - The research in this thesis has been done in the Bionanoscience for Membrane proteins group at the Department of Chemistry at University of Copenhagen under supervision of Assoc. Prof Karen L. Martinez. The main goal of the thesis has been to expand the scope of protein arrays to use 3D nanostructures in microfluidic devices for protein immobilization. Using a microfluidic approach allows us to perform hydrodyanmic flow focusing to address different locations on the microfluidic device to immobilize proteins on specific 3D nanostructures in a controlled manner. The flow focusing technique is the driving force to obtain protein multiplexing where different proteins are immobilzed on the same microfluidic device. Protein arrays are deemed fundamental for protein identification in high-throughput fashion used for diagnostics among other fields. Array platforms are extensively studied and engineered to increase their performance by higher sensitivity, lower detection limit and miniaturization. However problems such as limited sensitivity, critical detection limit and low signal-to-background ratio persist in conventional 2D protein arrays by spotting techniques. 3D structures such as vertical semiconducting nanowires and polymeric micropillars are interesting candidates to overcome these problems and are evaluated for protein immobilization in microfluidic devices. The first part of the thesis is theoretical calculations about the signal-to-background ratio of nanowires compared to micropillars followed by experiments to demonstrate protein immobilization on the structures monitored by fluorescence microscopy. The second part is the implementation of microfluidics where two devices are used to flow fluorescent biomolecule solution through the nanowire and micropillar samples respectively. The proof-of-concept of hydrodynamic flow focusing is reported and different fluorescent conjugates are immobilized on the micropillars obtaining protein multiplexing. The third part is the development and assessment of a kinase assay to detect endogenous phosphorylated ERK in cell lysates using nanowire samples to exploit their high signal-to-background ratio.

AB - The research in this thesis has been done in the Bionanoscience for Membrane proteins group at the Department of Chemistry at University of Copenhagen under supervision of Assoc. Prof Karen L. Martinez. The main goal of the thesis has been to expand the scope of protein arrays to use 3D nanostructures in microfluidic devices for protein immobilization. Using a microfluidic approach allows us to perform hydrodyanmic flow focusing to address different locations on the microfluidic device to immobilize proteins on specific 3D nanostructures in a controlled manner. The flow focusing technique is the driving force to obtain protein multiplexing where different proteins are immobilzed on the same microfluidic device. Protein arrays are deemed fundamental for protein identification in high-throughput fashion used for diagnostics among other fields. Array platforms are extensively studied and engineered to increase their performance by higher sensitivity, lower detection limit and miniaturization. However problems such as limited sensitivity, critical detection limit and low signal-to-background ratio persist in conventional 2D protein arrays by spotting techniques. 3D structures such as vertical semiconducting nanowires and polymeric micropillars are interesting candidates to overcome these problems and are evaluated for protein immobilization in microfluidic devices. The first part of the thesis is theoretical calculations about the signal-to-background ratio of nanowires compared to micropillars followed by experiments to demonstrate protein immobilization on the structures monitored by fluorescence microscopy. The second part is the implementation of microfluidics where two devices are used to flow fluorescent biomolecule solution through the nanowire and micropillar samples respectively. The proof-of-concept of hydrodynamic flow focusing is reported and different fluorescent conjugates are immobilized on the micropillars obtaining protein multiplexing. The third part is the development and assessment of a kinase assay to detect endogenous phosphorylated ERK in cell lysates using nanowire samples to exploit their high signal-to-background ratio.

UR - https://soeg.kb.dk/permalink/45KBDK_KGL/fbp0ps/alma99122361070005763

M3 - Ph.D. thesis

BT - A microfluidic approach for biological applications using vertical 3D nanostructures

ER -