Limitations of Microfluidic Emulsification

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Table of contents

Introduction
Colloids in Chapter 1 Biotechnology
1.1 Materials Science & Biotechnology
1.1.1 Colloidal Materials
1.2 Magnetic Particles
1.2.1 Structure & Properties
1.2.2 Magnetic Particle Applications
1.2.3 In Vivo Biomedical Applications
1.2.4 Microbeads & In Vitro Technologies
1.2.5 Examples of Magnetic Bead Based Technologies
1.3 Materials Perspective
1.3.1 Synthesis of Magnetic Particles
1.3.2 Particle Stability
1.3.1 Biofunctionalisation
1.3.2 Magnetic Particle Limitations
1.4 Liquid Particles
1.4.1 Types of Liquid Particle
1.4.2 Technological Applications
1.4.3 Biofunctionalisation
1.4.4 Magnetic Emulsions
1.5 Novel Magnetic Particles for Bioassays
Chapter 2 Emulsions & Formulation
2.1 Definition, Requirements & Types
2.1.1 Emulsion Stability
2.1.2 States of Emulsification
2.1.3 Project Implications
2.2 Surfactants & Stability
2.2.1 Surfactant Function
2.2.2 Surfactant Types
2.2.3 Phospholipids
2.2.4 Micelle Formation
2.2.5 Reversibility
2.2.6 The Bancroft Rule
2.2.1 Hydrophobic Lipophilic Balance
2.2.2 Pickering Emulsions
2.3 The Nature of the Dispersed Phase
2.3.1 Ferrofluids
2.4 Quantifying Emulsions
2.4.1 Volume Fraction
2.4.2 Mean Size & Size Distribution
2.4.3 Light Scattering
2.5 Creating an Emulsion
2.5.1 Emulsification Theory
2.5.2 Emulsification Techniques
2.6 Membrane & Microchannel Emulsification
2.6.1 Membrane Emulsification
2.6.2 Microchannel Emulsification
2.6.3 Microfluidic Chip Fabrication
2.6.1 Limitations of Microfluidic Emulsification
2.7 Emulsions & Formulation Conclusion
Chapter 3 Formulation Results
3.1 Introduction
3.2 Preparation of a Bulk Emulsion
3.2.1 Using a Cosurfactant
3.2.2 Preparation of a Bulk Emulsion
3.2.3 Emulsion Stability
3.2.4 Phospholipid Preparation
3.2.5 Visualisation of the Phospholipids
3.2.6 Poloxamer Stability Testing
3.2.7 Bulk Formulation Summary
3.3 Microfluidic Emulsification Results
3.3.1 Oil compatibility & Swelling
3.3.2 Surfactant Limits
3.3.3 Glass Chip
3.3.4 Calculating droplet concentration
3.3.5 Functional Droplets
3.3.6 Overall Microfluidic Formulation
3.4 The Ligand/Receptor Pair
3.4.1 Buffers & pH
3.5 Adding Ligands to the Interface
3.5.1 Measuring Protein Capture
3.5.2 Kinetics of Capture
3.5.3 Effects of PEG Spacer Length and Cosurfactant PEG Length
3.5.4 Quantifying Streptavidin Capture
3.5.5 Effect of Ligand Concentration
3.5.6 Ligands – Conclusion
3.6 Evidence of Fluidity
3.6.1 Adhesion Plaque Formation
3.6.2 Bead Capture & Mobility
3.6.3 Interfacial Fluidity Conclusion
3.7 Magnetic Content – Ferrofluid
3.7.1 Preparation of the ferrofluid
3.7.2 Ferrofluid Stability
3.7.3 Consequences of Aggregation
3.8 Formulation Results Summary
Chapter 4 Application – Agglutination Assay
4.1 Introduction
4.2 In Vitro Diagnostics & Particle Assays
4.2.1 Biomarkers
4.2.2 Bioassays & IVD
4.2.3 Targeting Biomarkers
4.2.4 Affinity Assays
4.2.5 Affinity Assay Limitations
4.3 Magnetic Particle Based Assays
4.3.1 Magnetic ELISA
4.3.2 Magnetic Agglutination:
4.3.3 Magnetic ELISA versus Magnetic Agglutination
4.3.4 Magnetic Particle Limitations for Agglutination
4.3.5 Emulsions as an Assay technology
4.4 Bulk Agglutination
4.4.1 Bulk Agglutination Conclusion
4.5 Flow Cytometry
4.5.1 The Flow Cytometer
4.5.2 Differentiation of population by size
4.5.3 Advantages of Flow Cytometry
4.5.4 Flow Cytometry Limitations
4.6 Flow Cytometry and Emulsions
4.6.1 Aggregation of the Bulk emulsion
4.6.2 Limitations of Measuring Emulsions
4.6.3 Conclusion
4.7 Measuring Agglutination using Flow Cytometry
4.7.1 3 μm M270 Streptavidin Beads
4.7.2 Reaction Protocol
4.7.3 Limit of Detection
4.7.4 1 μm Dynal MyOne Carboxy Streptavidin Beads
4.7.5 40 nm FS40 Neutravidin FluoSpheres
4.7.6 Streptavidin
4.7.7 Cytometric Agglutination Summary
4.8 Agglutination Conclusion
Chapter 5 Conclusion & Perspectives
5.1 Conclusion
5.1.1 Formulation & Production
5.1.2 Particle Application- Agglutination Assay
5.2 Perspectives
5.2.1 Agglutination measured by Flow Cytometry
5.2.1 Alternative Applications
Materials & Methods
Solubilisation of Phospholipids
Preparation of PDMS chips
5.3 Surface treatment of the PDMS chip
Microfluidic Emulsification Setup
Microscopy
Bibliography