Droplet generators are at the heart of many microfluidic devices developed for life science applications but are difficult to tailor to each specific application. The high fabrication costs, complex fluid dynamics, and incomplete understanding of multi-phase flows make engineering droplet-based platforms an iterative and resource-intensive process. First, we demonstrate the suitability of desktop micromills for low-cost rapid prototyping of thermoplastic microfluidic devices. With this method, microfluidic devices are made in 1 - 2 hours, have a minimum feature size of 75 μm, and cost less than $10. These devices are biocompatible and can accommodate integrated electrodes for sophisticated droplet manipulations, such as droplet sensing, sorting, and merging. Next, we leverage low-cost rapid prototyping to characterize the performance of microfluidic flow-focusing droplet generators. Specifically, the effect of eight design parameters on droplet diameter, generation rate, generation regime, and polydispersity are quantified. This was achieved through orthogonal design of experiments, a large-scale experimental dataset, and statistical analysis. Finally, we capitalize on the created dataset and machine learning to achieve accurate performance prediction and design automation of flow-focusing devices. The developed capabilities are captured in a software tool that converts high-level performance specifications to a device that delivers the desired droplet diameter and generation rate. This tool effectively eliminates the need for resource-intensive design iterations to achieve functional droplet generators. We also demonstrate the tool’s generalizability to new fluid combinations with transfer learning. We expect that our newly established framework on rapid prototyping, performance characterization informed by design of experiments, and machine learning guided design automation to enable extension to other microfluidic components and to facilitate widespread adoption of droplet microfluidics in the life sciences.

Thrombotic events are known to be associated with various cancers and recent research has implicated parts of the coagulation systemin promoting cancer progression. In particular, thrombin has been studied for its mitogenic effects in 2D cultures as well as in cancer progression in vivo in animal models however, conflicting results exist. Studies of proliferation in response to thrombin stimulation, of pancreatic cancer cells or pancreatic cancer-associated fibroblasts (CAFs) in vitro, that utilize a3D culture platform are significantly limited. In this study, PDAC cancer cells and cancer-associated fibroblast (CAF) cells were exposed to thrombin using a microfluidic device that mimics in vivo conditions. The cells used herein were cultured in a microfluid device, suspended inside of a 3D collagen matrix, and exposed to daily stimulation of 1 U/mL of thrombin in serum-free media for one hour. The findings of this study are that there is no statistically significant effect, promotive or inhibitory, on the proliferation of the cells used in this study, these results were unexpected. At the end of this paper, a review of potential reasons as to why no significant effect was seen on the cells is presented.

In this study the author reports the effects of surface roughness on dynamic behavior of water droplets on different types of rough structures. First, the influence of roughness geometry on the Wenzel/ Cassie-Baxter transition of water droplets on one-tier (solid substrates with Si micropillars) surfaces is studied (Chapter 3). In order to address distinct wetting behaviors of the advancing and receding motions, the author investigates the Wenzel/ Cassie-Baxter transition of water droplets on one-tier surfaces over a wide range of contact line velocities and droplet volumes in both advancing and receding movements. The discussions are strengthened by experimental results. According to the author’s analysis, the advancing contact zone tends to follow the Cassie-Baxter behavior for a wider range of geometric ratios than the receding contact zone. Physical phenomena such as advancing contact line rolling mechanism and the pinning of the receding contact line are introduced to justify distinct transition points of the advancing and receding movements respectively. Based on the analysis provided in Chapter 3, the author experimentally investigates the contact line fluctuations and contact line friction coefficients of water droplets on smooth, one-tier, and two-tier (with carbon nanotubes (CNTs) grown on Si micropillars) surfaces in Chapters 4 and 5. Both the advancing and receding contact line fluctuations/friction coefficients have been measured, analyzed and compared on smooth, one-tier, and two-tier surfaces over a wide range of contact line velocities and droplet volumes. A comprehensive analysis is provided to explain the experimental observations.

<p>Microfluidics has established itself as a key technology in a wide range of fields including pharmaceuticals, point-of-care diagnostics, thermal management, and space technology. Most of these applications involve manipulation of small quantities (micro – nanoliters) of fluids and various particles or biological cells suspended in them. These platforms employ mechanical, thermal, acoustic, magnetic, optical, electric and many other means for creating particle and fluid motion. Many biological applications require handling cells that are vulnerable to getting damaged if proper physiological conditions are not maintained or if excessive force is applied on them. The non-invasive nature of optical and electrical micro-manipulation techniques such as rapid electrokinetic patterning (REP) has proven to be of great importance in such applications. These techniques enable handling, transportation, sorting and arrangement of fragile synthetic micro/nanoparticles and biological cells without compromising their structure and surface properties.</p> <p>REP is a recently developed micro-manipulation tool that employs optically induced electrothermal vortices to create custom flow patterns. Particle suspensions are entrained in these vortices and are trapped on an electrode surface through AC electrokinetic mechanisms. This work focuses on characterizing a REP trap and discusses its potential applications in handling biological cells. Polystyrene microparticles are confined in a REP trap and a MATLAB program is used to track their motion inside the trap. The tracked particle trajectory reveals that the potential energy of the trapped particle is parabolic and hence the trap is Hookean in nature. The trap is modelled as a spring-mass system and the stiffness coefficient of that system is found to be of the order of 10^-15 N/μm. The origin of the restoring force in the spring-mass model is found to be the drag force created by the electrothermal vortex. The ability to exert ultra-small forces in a stable trap enables REP to be used in various non-invasive particle manipulation applications.</p> <p>The transient nature of REP is studied using numerical modeling and particle image velocimetry (PIV) analysis of a vortex created by a moving laser spot. A numerical model suggests that custom-shaped steady state REP vortices can be created via superposition of multiple axisymmetric circular shaped vortices. However, the method of superposition cannot be extended to transient traps and a more involved 3D model is required to simulate them. The laser spot is scanned back-and-forth in a line with different speeds to create transient REP vortices. The PIV analysis, in agreement with the numerical model, shows that the location of the moving vortex is undiscernible at high speeds. Moreover, the circular shaped vortex is stretched out into a line when the laser scanning frequencies are more than 15 Hz.</p> <p>The particle-electrode attraction force, which entraps the particles at the electrode surface, is characterized using particle diffusometry (PD) and defocusing particle tracking. PD is used to measure the diffusion coefficient of polystyrene particles under different electric field parameters near an electrode surface. It is found that the particle diffusivity decreases with a decrease in the electric field frequency from 150 – 30 kHz and with an increase in the applied voltage from 4 – 8 V_pp. A MATLAB program is used to track the number of in-focus particles and their distance from the electrode surface. A histogram of the particles’ distance from the electrode surface shows an increase in the particle concentration near the electrode at low frequencies (30 – 60 kHz). These observations suggest that the average height of an entrapped particle decreases with a decrease in applied field frequency and an increase in applied voltage. This suggests that the attractive trapping force is significant at 30 kHz but diminishes at around 150 kHz.</p> <p>Salt and sugar-based isotonic media used for cell suspensions pose several challenges for electrokinetic mechanisms such as REP. Various solutions to overcome these challenges for bio-manipulation applications are discussed in this work. The presence of DC offset in the AC electric field is found to enhance particle entrapment in sugar-based media. The effect of DC offset on trapping performance in bio-relevant media is assessed by measuring the stability of the REP trap. This work also shows entrapment and manipulation of Mice pancreatic cancer cells (KPC2) suspended in the sugar-based isotonic media using REP. The biological applications of the REP technology are highly promising, but they have not yet been well-explored. This work lays the foundation of understanding how REP can be operated in high osmolarity media for bio-manipulation applications.</p>

This thesis details the design, fabrication, and testing of two optofluidic platforms, a square fused silica capillary and a MgF2-PDMS microfluidic chip to detect radiation-induced biochemical changes in cells during radiation treatment (radiotherapy). The platforms integrate a near-infrared Raman system of 785 nm excitation and a fiber-based optical trap at 1064 nm in a dual-beam configuration for the manipulation and subsequent examination of single polystyrene beads (5µm) and two breast carcinoma cell lines, MCF-7, and MDA-MB-23 (20-30 µm). Particular attention was paid to the role of MgF2 as a novel substrate for microfluidic fabrication and the device background contributions that could hinder spectral contributions from the samples. Successful optical trapping within the platforms was performed, which allowed the sample immobilization for the entire Raman acquisition time (10-30 s) via an orthogonally positioned objective for the excitation and collection of Raman signal. Data collected in the MgF2-PDMS microchip yielded high-quality spectra with no presence of PDMS characteristic Raman peaks in the spectral region of 450-1800 cm-1. / Graduate / 2023-04-08

Separation of 225Ra (t1/2 = 15 d) from its daughter isotope 225Ac (t1/2 = 10 d) is necessary to obtain pure 225Ac for cancer alpha-therapy. In this study, microscale separation of 225Ra from its daughter 225Ac using BioRad AG50X4 cation exchange resin was achieved with good reproducibility across microdevices, and ≥90% purity was achieved for 225Ac, which is comparable to conventional chromatography. These results indicate the potential for greater use of microfluidics for biomedical radiochemistry. The modularity of the system and its compatibility with different resins allows for quick and easy adaptation to the various needs of a separation campaign.

Primary open-angle glaucoma (POAG) is a leading cause of blindness in the world. A primary risk factor for POAG is elevated intraocular pressure (IOP), caused by increased aqueous humor outflow resistance. Currently, lowering the IOP is the only effective way of treating glaucoma; however, the cause of increased outflow resistance remains unclear. This thesis will present a series of studies which investigated structures of the trabecular outflow pathway, including Schlemm’s canal endothelium, juxtacanalicular tissue, and trabecular beams, and their roles in regulating aqueous outflow resistance. The studies were conducted in both human and animal models using ex vivo ocular perfusion as well as in vitro microfluidic systems. In the first study, we investigated the effects of Y27632, a derivative of Rho-kinase inhibitor that is being developed as next generation glaucoma drug with unclear IOP lowering mechanism, on aqueous humor outflow dynamics and associated morphological changes in normal human eyes and laser-induced ocular hypertensive monkey eyes. In the second study, we developed and validated a novel three-dimensional microfluidic system using lymphatic microvascular endothelial cells. The microfluidic system can be used to study Schlemm’s canal endothelial cell dynamics and aqueous humor transport mechanism in the future. In the last study, we characterized the morphological structure, distribution, and thickness of the endothelial glycocalyx in the aqueous humor outflow pathway of human and bovine eyes. Together these studies will help define new directions for therapy that will help control IOP and preserve vision throughout a normal life span.

Polymer nanoparticles (PNPs) that exhibit selective stimuli-responsive degradation and drug release at tumor sites are promising candidates in the development of smart nanomedicines. In this thesis, we demonstrate a microfluidic approach to manufacturing biological stimuli-responsive PNPs with flow-tunable physicochemical and pharmacological properties. The investigated PNPs contain cleavable disulfide linkages in two different locations (core and interface, DualM PNPs) exhibiting responsivity to elevated levels of glutathione (GSH), such as those found within cancerous cells. First, we conduct a mechanistic study on the microfluidic formation of DualM PNPs without encapsulated drug. We show that physicochemical properties, including size, morphology, and internal structure, of DualM PNPs are tunable with manufacturing flow rate. Microfluidic formation of DualM PNPs is explained by the interplay of shear-induced coalescence, shear-induced breakup, and intraparticle chain rearrangements. In addition, we demonstrate that rates of GSH-triggered changes in size and internal structure are linearly correlated with initial PNP sizes and internal structures, respectively. Next, we expand our study to focus on microfluidic control of pharmacological properties of DualM PNPs containing either an anticancer drug (paclitaxel, PAX-PNPs) or a fluorescent drug surrogate (DiI-PNPs). Microfluidic PAX-PNPs and DiI-PNPs show similar sizes and morphologies with their non-drug-loaded counterparts under the same flow conditions. We then show that pharmacological properties of DualM PNPs, including encapsulation efficiency, GSH-triggered release rate, cell uptake, cytotoxicity against MCF-7 (cancerous) and HaCaT (healthy), and relative difference in MCF-7 and HaCaT cytotoxicity, all increase linearly as flow-directed PNP size decreases, providing remarkably simple process-structure-property relationships. In addition, we show that microfluidic manufacturing improves encapsulation homogeneities within PNPs relative to bulk nanoprecipitation. These results highlight the potential of flow-directed shear processing in microfluidics for providing controlled manufacturing routes to biological stimuli-responsive nanomedicines optimized for specific therapeutic applications. Finally, we summarize various design strategies of biological stimuli-responsive PNPs. We show that the location and density of disulfide linkages within PNPs determines stimulus-triggered degradation mechanism and kinetics. In addition, we show various bottom-up approaches to tune PNP responsivities that involves chemical processing, including formulation chemistry and intramolecular forces. Along with the top-down microfluidic approach that we demonstrate experimentally, this chapter provides a more comprehensive understanding of process-structure-property relations opening up vast possibilities for manufacturing smarter nanomedicines. / Graduate

Les tests « in vitro » permettent d’établir près de 70% des diagnostics et leur développement pour une utilisation au plus près du patient apparaît donc comme un enjeu majeur de santé publique. Dans ce contexte, les critères ASSURED (« Affordable, Sensitive, Specific, User-friendly, Rapid and robust, Equipment-free and Deliverable to end-users ») a été défini par l’organisation mondiale de la santé pour que les chercheurs développent des outils de diagnostic dits « Point of Care » utilisables par le plus grand nombre. Avec l’essor de la microfluidique, la gamme des dispositifs possibles s'est élargie et des biocapteurs intégrés ont été développés, transformant le signal biologique d’une reconnaissance d’un biomarqueur par une sonde biologique en un signal optique, électrochimique, mécanique ou encore magnétique. Comme les milieux biologiques sont en grande majorité amagnétiques, les capteurs magnétiques ne sont pas affectés par l’utilisation de matrices biologiques complexes comme peuvent l’être les mesures optiques ou électrochimiques. De plus ces capteurs sont faciles à produire et intégrables dans les puces microfluidiques. Cette thèse a pour objectifs de concevoir un outil de diagnostic in vitro basé sur des capteurs à magnétorésistance géante et de tester ses performances. Cette étude a été réalisée en utilisant une lignée cellulaire de myélome murin. Les cellules sont marquées spécifiquement par des particules magnétiques fonctionnalisées par des anticorps dirigés contre un de leurs antigènes et sont passées dans le canal microfluidique au-dessus des capteurs. Cette méthode de détection dynamique permet de compter les objets magnétiques un par un. La difficulté réside dans la distinction des signaux spécifiques provenant des cellules marquées des signaux faux positifs induits par les billes restant en solution. Deux types de dispositifs ont été conçus dans cette thèse pour lever ce verrou. Le premier possède une couche inerte de séparation de quelques micromètres entre les capteurs GMR et le canal qui permet de supprimer les signaux des billes isolées. Le second dispositif, qui a des capteurs à la fois au-dessus et au-dessous du canal microfluidique, permet une double détection simultanée de chaque objet magnétique. Il est ainsi possible de connaître le nombre de billes qui les marquent et de déterminer s’il s’agit d’un agrégat de billes ou d’un objet biologique. / The « in vitro » tests are requested for the establishment of nearly 70% of diagnoses and their development for on-site detection therefore appears to be a major public health issue. In this context, the ASSURED criterion (« Affordable, Sensitive, Specific, User-friendly, User-friendly, Rapid and robust, Equipment-free and Deliverable to end-users ») has been defined by the World Health Organization to encourage researchers to develop diagnostic tools called « Point of Care » that can be widely used.With the rise of microfluidics, the range of possible devices has broadened and integrated biosensors have been developed, transforming the biological signal from a biomarker recognition by a biological probe into an optical, electrochemical, mechanical or magnetic signal. As biological environments are largely non-magnetic, magnetic sensors are not affected by the use of complex biological matrices as are optical or electrochemical measurements. In addition, these sensors are easy to produce and can be integrated into microfluidic chips. The objectives of this thesis are to design a diagnostic tool in vitro based on giant magnetoresistance sensors and to test its performance. Its development was carried out using a murine myeloma cell line. The cells are specifically labeled by magnetic particles functionalized by antibodies directed against one of their antigens and flown in the microfluidic channel above the sensors. This dynamic detection method allows magnetic objects to be counted one by one. The challenge is to distinguish the signals coming from the labeled cells from those of the beads remaining in solution. In order to address this problem, two labs on chips are developed in this thesis. In a first device, an inner layer of a few micrometers separates the sensors from the channel which allows to suppress the signals of the isolated beads. The second device has sensors both above and below the microfluidic channel and can measure the number of beads corresponding to each doubly detected object which can thus be identified (aggregates or biological objects).

Nous nous sommes intéressés à l'auto-enroulement de films de polydimethylsiloxane (PDMS) oxydés dans des vapeurs de solvant. Brièvement, des films minces de PDMS sont obtenus par enduction sous centrifugation. Ces films sont ensuite exposés à un plasma d'oxygène, ce qui a pour conséquence d'oxyder et de rigidifier leurs surfaces. Lorsque ces systèmes sont exposés à certains solvants en phase gazeuse, le PDMS non-oxydé gonfle. Cela mène à l'auto-enroulement des films et donc à la formation de capillaires. Ce mécanisme est intéressant pour la fabrication de canaux microfluidiques car ce qui deviendra la surface interne desdits canaux peut-être caractérisé et fonctionalisé avant l'enroulement.Dans un premier chapitre, différents aspects de l'auto-enroulement sont passés en revue théoriquement et numériquement.Un second chapitre expérimental est dédié à l'étude de la couche oxydée par nano-indentation AFM. Les propriétés mécaniques du système composite (couche dur sur substrat mou) sont mesurées et interprétées au moyen d'un nouveau modèle pour extraire notamment l'épaisseur du film oxydé.Dans un troisième chapitre, l'auto-enroulement des tubes lui-même est étudié. Le diamètre interne des capillaires obtenus en fonction de paramètres expérimentaux est examiné et confronté à la théorie. Plusieurs démonstrations de principe de tube avec une surface interne fonctionnalisée sont fournies.Enfin, pour répondre à des problématiques d'intégration des systèmes dans une structure microfluidique plus complexe, une méthode innovante est proposée dans un quatrième et dernier chapitre. Basée sur l'impression jet d'encre de moules sacrificiels, la méthode est d'abord mise en place expérimentalement. De nombreuses démonstrations de principe du vaste potentiel de cette idée sont ensuites proposées. / The guideline of this work is the spontaneous rolling of oxidized polydimethylsiloxane (PDMS) thin films in organic solvant vapors. Briefly, thin films of PDMS are produced by spin coating. Those films are then exposed to oxygen plasma which oxidizes and hardens their surfaces. When those systems are immersed in appropriate solvent vapors, non oxidized PDMS selectively swells. This leads to the spontaneous rolling of the films and thus to the formation of capillaries. This mechanism is of great interest for the fabrication of microfluidic channels because what is to become the inner surface of those channels can be characterized and functionalized prior to rolling.In a first chapter, different aspects of spontaneous rolling are reviewed theoretically and numerically.A second chapter is dedicated to the investigation of the oxide layer by AFM nanoindentation. The mechanical properties of the composite system (hard layer on a soft substrate) are measured and interpreted with a new model in order to extract in particular the thickness of the oxide layer.A third chapter dwells on engineering of the rolled-up tubes. The inner diameter of the capillaries as a function of experimental parameters is measured and confronted to theory. We present tubes with various inner surface functionalizations as a proof of concept of the method.Finally, in order to solve the issue of the integration of the system in a wider structure, an innovative method is proposed in a final fourth chapter. Based on the fabrication of a sacrificial mold by inkjet printing, the method is first established and implemented. Several proof-of-concept systems are then displayed in order to demonstrate the great potential of that idea.