Custom-Designed Biohybrid Micromotor for Potential Disease Treatment

Micromotors are recognized as promising candidates for untethered micromanipulation and targeted cargo transport. Their future application is, however, hindered by the low efficiency of drug encapsulation and their poor adaptability in physiological conditions. To address these challenges, one potential solution is to incorporate micromotors with biological materials as the combination of functional biological entities and smart artificial parts represents a manipulable and biologically friendly approach. This dissertation focuses on the development of custom-designed micromotors combined with sperm and their potential applications on targeted diseases treatment. By means of 2D and 3D lithography methods, microstructures with complex configurations can be fabricated for specific demands. Bovine and human sperm are both for the first time explored as drug carriers thanks to their high encapsulation efficiency of hydrophilic drugs, their powerful self-propulsion and their improved drug-uptake relying on the somatic-cell fusion ability. The hybrid micromotors containing drug loaded sperm and constructed artificial enhancements can be self-propelled by the sperm flagella and remotely guided and released to the target at high precision by employing weak external magnetic fields. As a result, micromotors based on both bovine and human sperm show significant anticancer effect. The application here can be further broadened to other biological environments, in particular to the blood stream, showing the potential on the treatment of blood diseases like blood clotting. Finally, to enhance the treatment efficiency, in particular to control sperm number and drug dose, three strategies are demonstrated to transport swarms of sperm. This research paves the way for the precision medicine based on engineered sperm-based micromotors

The Role of Hydrophilic Sandblasted Titanium and Laser Microgrooved Zirconia Surfaces in Dental Implant Treatment

Dental implant surface modifications affect surface roughness, chemistry, topography, and consequently influence biological bone response. Current surface treatments are directed toward increased hydrophilicity and wettability of dental surfaces that allow earlier implant loading due to accelerated osseointegration. This is clinically reflected in increased implant stability and mainteined crestal bone level. Further modification includes microgrooving of zirconia implants by femtosecond laser ablation. Favorable initial results encourage further clinical assessment of this microgrooved zirconia implants

Intracellular delivery by membrane disruption: Mechanisms, strategies, and concepts

© 2018 American Chemical Society. Intracellular delivery is a key step in biological research and has enabled decades of biomedical discoveries. It is also becoming increasingly important in industrial and medical applications ranging from biomanufacture to cell-based therapies. Here, we review techniques for membrane disruption-based intracellular delivery from 1911 until the present. These methods achieve rapid, direct, and universal delivery of almost any cargo molecule or material that can be dispersed in solution. We start by covering the motivations for intracellular delivery and the challenges associated with the different cargo typesñYsmall molecules, proteins/peptides, nucleic acids, synthetic nanomaterials, and large cargo. The review then presents a broad comparison of delivery strategies followed by an analysis of membrane disruption mechanisms and the biology of the cell response. We cover mechanical, electrical, thermal, optical, and chemical strategies of membrane disruption with a particular emphasis on their applications and challenges to implementation. Throughout, we highlight specific mechanisms of membrane disruption and suggest areas in need of further experimentation. We hope the concepts discussed in our review inspire scientists and engineers with further ideas to improve intracellular delivery

A fixed-target platform for serial femtosecond crystallography in a hydrated environment.

For serial femtosecond crystallography at X-ray free-electron lasers, which entails collection of single-pulse diffraction patterns from a constantly refreshed supply of microcrystalline sample, delivery of the sample into the X-ray beam path while maintaining low background remains a technical challenge for some experiments, especially where this methodology is applied to relatively low-ordered samples or those difficult to purify and crystallize in large quantities. This work demonstrates a scheme to encapsulate biological samples using polymer thin films and graphene to maintain sample hydration in vacuum conditions. The encapsulated sample is delivered into the X-ray beam on fixed targets for rapid scanning using the Roadrunner fixed-target system towards a long-term goal of low-background measurements on weakly diffracting samples. As a proof of principle, we used microcrystals of the 24 kDa rapid encystment protein (REP24) to provide a benchmark for polymer/graphene sandwich performance. The REP24 microcrystal unit cell obtained from our sandwiched in-vacuum sample was consistent with previously established unit-cell parameters and with those measured by us without encapsulation in humidified helium, indicating that the platform is robust against evaporative losses. While significant scattering from water was observed because of the sample-deposition method, the polymer/graphene sandwich itself was shown to contribute minimally to background scattering

Cavitation in soft matter

Cavitation is the sudden, unstable expansion of a void or bubble within a liquid or solid subjected to a negative hydrostatic stress. Cavitation rheology is a field emerging from the development of a suite of materials characterization, damage quantification, and therapeutic techniques that exploit the physical principles of cavitation. Cavitation rheology is inherently complex and broad in scope with wide-ranging applications in the biology, chemistry, materials, and mechanics communities. This perspective aims to drive collaboration among these communities and guide discussion by defining a common core of high-priority goals while highlighting emerging opportunities in the field of cavitation rheology. A brief overview of the mechanics and dynamics of cavitation in soft matter is presented. This overview is followed by a discussion of the overarching goals of cavitation rheology and an overview of common experimental techniques. The larger unmet needs and challenges of cavitation in soft matter are then presented alongside specific opportunities for researchers from different disciplines to contribute to the field

Femtosecond laser microfabricated devices for biophotonic applications

Femtosecond Laser DirectWriting has emerged as a key enabling technology for realising miniaturised biophotonic applications offering clear advantages over competing soft-lithography, ion-exchange and sol-gel based fabrication techniques. Waveguide writing and selective etching with three-dimensional design flexibility allows the development of innovative and unprecedented optofluidic architectures using this technology. The work embodied in this thesis focuses on utilising the advantages offered by direct laser writing in fabricating integrated miniaturised devices tailored for biological analysis. The first application presented customised the selective etching phenomenon in fused silica by tailoring the femtosecond pulse properties during the writing process. A device with an embedded network of microchannels with a significant difference in aspect-ratio was fabricated, which was subsequently applied in achieving the high-throughput label-free sorting of mammalian cells based on cytoskeletal deformability. Analysis on the device output cell population revealed minimal effect of the device on cell viability. The second application incorporated an embedded microchannel in fused silica with a monolithically integrated near-infrared optical waveguide. This optofluidic device implemented the thermally sensitive emission spectrum of semiconductor nanocrystals in undertaking remote thermometry of the localised microchannel environment illuminated by the waveguide. Aspects relating to changing the wavelength of illumination from the waveguide were analysed. The effect of incorporating carbon nanotubes as efficient heaters within the microchannel was investigated. Spatio-thermal imaging of the microchannel illuminated by the waveguide revealed the thermal effects to extend over distances appreciably longer than the waveguide cross-section. On the material side of direct laser writing, ultra-high selective etching is demonstrated in the well-known laser crystal Nd:YAG. This work presents Nd:YAG as a material with the potential to develop next-generation optofluidic devices

Femtosecond Laser Nanomachining and Applications to Micro/Nanofluidics for Single Cell Analysis.

Femtosecond laser machining has huge potential to impact micro/nanofluidics with its ability to arbitrarily abricate 3-dimensional geometries with feature sizes down to nanometer scales. Because cleanroom facilities, multilayer configurations, and glass bonding are not necessary to achieve 3-dimensional subsurface nanofeatures in glass, current planar lithography-etch-bond processes are easily combined with femtosecond laser machining; a hybrid machining based on these two methods constitutes a promising fabrication method for next generation microchip processes. The major challenge facing fs laser machining is that increasing the length of subsurface capillaries is very difficult; the normalized length (length/diameter) had previously been limited to 50. In this dissertation, a new phenomenon, acoustic nodeformation, is shown to be the major barrier to increasing capillary length, and a theoretical model for node formation is established. Based on the node equation, degassed water, which is introduced to the ablation site to assist machining, is found to substantially overcome node formation. Thus, a novel degassed-water-assisted fs laser machining process is developed, improving the normalized length of submicron-scale capillaries to longer than 1000. Nano-capillary electrophoresis (nCE) is demonstrated, initiating a submicronscale separation regime with millisecond-fast separations and 1 femtoliter injection volumes (1000 times smaller than a single cell volume). Also, the current-controlled dielectric breakdown is found to convert a thin glass wall to an electrode, which is the core part in the nCE device zero-flow sample loader. This phenomenon can be further exploited in many novel micro/nanofluidic modules such as electrokinetic pumps, nanosensors, and nanoactuators with freedom to directly embed these modules in glass chips. These new micro/nanofluidic devices and modules will contribute to many novel biotechnology investigations, including single cell proteomics, cell characterization, DNA analysis, electrophysiology, and biological assays.Ph.D.Mechanical EngineeringUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/58515/1/toshlee_1.pd

Computationally efficient methods for modelling laser wakefield acceleration in the blowout regime

Electron self-injection and acceleration until dephasing in the blowout regime is studied for a set of initial conditions typical of recent experiments with 100 terawatt-class lasers. Two different approaches to computationally efficient, fully explicit, three-dimensional particle-in-cell modelling are examined. First, the Cartesian code VORPAL using a perfect-dispersion electromagnetic solver precisely describes the laser pulse and bubble dynamics, taking advantage of coarser resolution in the propagation direction, with a proportionally larger time step. Using third-order splines for macroparticles helps suppress the sampling noise while keeping the usage of computational resources modest. The second way to reduce the simulation load is using reduced-geometry codes. In our case, the quasi-cylindrical code CALDER-CIRC uses decomposition of fields and currents into a set of poloidal modes, while the macroparticles move in the Cartesian 3D space. Cylindrical symmetry of the interaction allows using just two modes, reducing the computational load to roughly that of a planar Cartesian simulation while preserving the 3D nature of the interaction. This significant economy of resources allows using fine resolution in the direction of propagation and a small time step, making numerical dispersion vanishingly small, together with a large number of particles per cell, enabling good particle statistics. Quantitative agreement of the two simulations indicates that they are free of numerical artefacts. Both approaches thus retrieve physically correct evolution of the plasma bubble, recovering the intrinsic connection of electron self-injection to the nonlinear optical evolution of the driver

Advanced optical systems for imaging and fabrication

Advanced optical systems for imaging and fabricatio

Three dimensional optofluidic devices for manipulation of particles and cells

Optical forces offer a powerful tool for manipulating single cells noninvasively. Integration of optical functions within microfluidic devices provides a new freedom for manipulating and studying biological samples at the micro scale. In the pursuit to realise such microfluidic devices with integrated optical components, Ultrafast Laser Inscription (ULI) fabrication technology shows great potential. The uniqueness and versatility of the technique in rapid prototyping of 3D complex microfluidic and optical elements as well as the ability to perform one step integration of these elements provides exciting opportunities in fabricating novel devices for biophotonics applications. The work described in this thesis details the development of three dimensional optofluidic devices that can be used for biophotonics applications, in particular for performing cell manipulation and particle separation. Firstly, the potential of optical forces to manipulate cells and particles in ULI microfluidic channels is investigated. The ability to controllably displace particles within a ULI microchannel using a waveguide positioned orthogonal to it is explored in detail. We then prototype a more complex 3D device with multiple functionalities in which a 3D optofluidic device containing a complex microchannel network and waveguides was used for further investigations into optical manipulation and particle separation. The microfluidic channel network and the waveguides within the device possess the capability to manipulate the injected sample fluid through hydrodynamic focusing and optically manipulate the individual particles, respectively. This geometry provided a more efficient way of investigating optical manipulation within the device. Finally, work towards developing a fully optimised 3D cell separator device is presented. Initial functional validation was performed by investigating the capability of the device to route particles through different outlet channels using optical forces. A proof of concept study demonstrates the potential of the device to use for cell separation based on the size of the cells. It was shown that both passive and active cell separation is possible using this device