Microfabricated platforms to investigate cell mechanical properties

Mechanical stimulation has been imposed on living cells using several approaches. Most early investigations were conducted on groups of cells, utilizing techniques such as substrate deformation and flow-induced shear. To investigate the properties of cells individually, many conventional techniques were utilized, such as AFM, optical traps/optical tweezers, magnetic beads, and micropipette aspiration. In specific mechanical interrogations, microelectro- mechanical systems (MEMS) have been designed to probe single cells in different interrogation modes. To exert loads on the cells, these devices often comprise piezo-electric driven actuators that attach directly to the cell or move a structure on which cells are attached. Uniaxial and biaxial pullers, micropillars, and cantilever beams are examples of MEMS devices. In this review, the methodologies to analyze single cell activity under external loads using microfabricated devices will be examined. We will focus on the mechanical interrogation in three different regimes: compression, traction, and tension, and discuss different microfabricated platforms designed for these purposes

Recursive Least Squares Filtering Algorithms for On-Line Viscoelastic Characterization of Biosamples

The mechanical characterization of biological samples is a fundamental issue in biology and related fields, such as tissue and cell mechanics, regenerative medicine and diagnosis of diseases. In this paper, a novel approach for the identification of the stiffness and damping coefficients of biosamples is introduced. According to the proposed method, a MEMS-based microgripper in operational condition is used as a measurement tool. The mechanical model describing the dynamics of the gripper-sample system considers the pseudo-rigid body model for the microgripper, and the Kelvin–Voigt constitutive law of viscoelasticity for the sample. Then, two algorithms based on recursive least square (RLS) methods are implemented for the estimation of the mechanical coefficients, that are the forgetting factor based RLS and the normalised gradient based RLS algorithms. Numerical simulations are performed to verify the effectiveness of the proposed approach. Results confirm the feasibility of the method that enables the ability to perform simultaneously two tasks: sample manipulation and parameters identification

A microgripper for single cell manipulation

This thesis presents the development of an electrothermally actuated microgripper for the manipulation of cells and other biological particles. The microgripper has been fabricated using a combination of surface and bulk micromachining techniques in a three mask process. All of the fabrication details have been chosen to enable a tri-layer, polymer (SU8) - metal (Au) - polymer (SU8), membrane to be released from the substrate stress free and without the need for sacrificial layers. An actuator design, which completely eliminates the parasitic resistance of the cold arm, is presented. When compared to standard U-shaped actuators, it improves the thermal efficiency threefold. This enables larger displacements at lower voltages and temperatures. The microgripper is demonstrated in three different configurations: normally open mode, normally closed mode, and normally open/closed mode. It has-been modelled using two coupled analytical models - electrothermal and thermomechanical - which have been custom developed for this application. Unlike previously reported models, the electrothermal model presented here includes the heat exchange between hot and cold arms of the actuators that are separated by a small air gap. A detailed electrothermomechanical characterisation of selected devices has permitted the validation of the models (also performed using finite element analysis) and the assessment of device performance. The device testing includes electrical, deflection, and temperature measurements using infrared (IR) thermography, its use in polymeric actuators reported here for the first time. Successful manipulation experiments have been conducted in both air and liquid environments. Manipulation of live cells (mice oocytes) in a standard biomanipulation station has validated the microgripper as a complementary and unique tool for the single cell experiments that are to be conducted by future generations of biologists in the areas of human reproduction and stem cell research

NEMS by sidewall transfer lithography

A batch fabrication process for nano-electro-mechanical systems (NEMS) based on sidewall transfer lithography (STL) is developed and demonstrated. The STL is used to form nanoscale flexible silicon suspensions entirely by conventional lithography. A two-step process is designed for single-layer STL to fabricate simple electrothermal actuators, while a three-step process is designed to allow nanoscale features intersecting with each other for more complicated device lay-outs. Fabricated nanoscale features has a minimum in-plane width of approx. 100nm and a high aspect ratio of 50 : 1. Combined structures with microscale and nanoscale parts are transferred together into silicon by deep reactive etching (DRIE). Suspensions are achieved either by plasma undercut or HF vapour etch based on BSOI. The STL processes are used to form nanoscale suspensions while conventional lithography is used to form localised microscale features such as anchors. A wide variety of demonstrator devices have been fabricated with high feature quality. Analytic models have been developed to compare with experimental characterization and finite element analysis (FEA) predictions. Lattice structures fabricated by multi-layer STL have also be investigated as a novel type of mechanical metamaterial. Thus, the process could allow low-cost and mass parallel fabrication of future NEMS with a wider range of potential applications.Open Acces

Concurrent Spatial Mapping of the Viscoelastic Behavior of Heterogeneous Soft Materials Via a Polymer-Based Microfluidic Device

This dissertation presents a novel experimental technique, namely concurrent spatial mapping (CSM), for measuring the viscoelastic behavior of heterogeneous soft materials via a polymer-based microfluidic device. Comprised of a compliant polymer microstructure and an array of electrolyte-enabled distributed resistive transducers, the microfluidic device detects both static and dynamic distributed loads. Distributed loads deform the polymer microstructure and are recorded as resistance changes at the locations of the transducers. The CSM technique identifies the elastic modulus of soft materials by applying a precisely controlled indentation depth using a rigid probe to a sample placed on the device. The spatially-varying elastic modulus of the sample translates to a non-uniform load, causing a non-uniform deformation of the microstructure and variations in the recorded resistance changes. The CSM technique measures the loss modulus of soft materials through a dynamic measurement by applying varying sinusoidal loads to a sample placed on the device. The spatially-varying loss modulus of the sample causes the microstructure to respond with corresponding time delay. Consequently, the phase shift between the sinusoidal load and deflection of the sample along its length are captured by the distributed transducers. As the first step of the experimental protocol, control experiments are implemented on the device to determine its static performance and system-level dynamic parameters. Next, the CSM technique is applied to both homogeneous and heterogeneous synthetic soft materials to measure their elastic moduli by applying a precisely controlled indentation depth through a probe, and the recorded load and device deflection are the output. The data are processed to obtain the overall load and the deflection of the sample at each transducer location and are further used to extract the elastic modulus distribution of the sample. The CSM technique is then applied to measure the loss modulus of soft materials. The measurable sinusoidal loads are the input, and the sinusoidal deflections of the device are the output. By applying the Fast Fourier Transform (FFT) algorithm and the nonlinear regression method, the data are processed to obtain the phase shift between the applied load and the device response along its microchannel length as well as the system-level parameters, namely stiffness (K), damping coefficient (D), and mass (M). In conjunction with the system-level parameters of the system with the device, obtained from the control experiment, the stiffness and the damping coefficient of a sample are calculated, and the sample’s loss modulus distribution is estimated accordingly. This CSM technique successfully measures the spatially-varying elastic modulus and loss modulus of soft materials. As compared with the nanoindentation-based technique, the CSM technique demonstrates its efficiency in spatially mapping the viscoelastic behavior of a sample without excluding interactions among neighboring compositions in a sample

Coupled finite element-finite volume multi-physics analysis of MEMS electrothermal actuators

Microelectromechanical systems (MEMS) are the instruments of choice for high-precision manipulation and sensing processes at the microscale. They are, therefore, a subject of interest in many leading industrial and academic research sectors owing to their superior potential in applications requiring extreme precision, as well as in their use as a scalable device. Certain applications tend to require a MEMS device to function with low operational temperatures, as well as within fully immersed conditions in various media and with different flow parameters. This study made use of a V-shaped electrothermal actuator to demonstrate a novel, state-of-the-art numerical methodology with a two-way coupled analysis. This methodology included the effects of fluid–structure interaction between the MEMS device and its surrounding fluid and may be used by MEMS design engineers and analysts at the design stages of their devices for a more robust product. Throughout this study, a thermal–electric finite element model was strongly coupled to a finite volume model to incorporate the spatially varying cooling effects of the surrounding fluid (still air) onto the V-shaped electrothermal device during steady-state operation. The methodology was compared to already established and accepted analysis methods for MEMS electrothermal actuators in still air. The maximum device temperatures for input voltages ranging from 0 V to 10 V were assessed. During the postprocessing routine of the two-way electrothermal actuator coupled analysis, a spatially-varying heat transfer coefficient was evident, the magnitude of which was orders of magnitude larger than what is typically applied to macro-objects operating in similar environmental conditions. The latter phenomenon was correlated with similar findings in the literature.peer-reviewe

Computer Vision Measurements for Automated Microrobotic Paper Fiber Studies

The mechanical characterization of paper fibers and paper fiber bonds determines the key parameters affecting the mechanical properties of paper. Although bulk measurements from test sheets can give average values, they do not yield any real fiber-level data. The current, state-of-the-art methods for fiberlevel measurements are slow and laborious, requiring delicate manual handling of microscopic samples. There are commercial microrobotic actuators that allow automated or tele-operated manipulation of microscopic objects such as fibers, but it is challenging to acquire the data needed to guide such demanding manipulation. This thesis presents a solution to the illumination problem and computer vision algorithms for obtaining the required data. The solutions are designed for a microrobotic platform that comprises actuators for manipulating the fibers and one or two microscope cameras for visual feedback.The algorithms have been developed both for wet fibers, which can be treated as 2D objects, and for dry fibers and fiber bonds, which are treated as 3D objects. The major innovations in the algorithms are the rules for the micromanipulation of the curly fiber strands and the automated 3D measurements of microscale objects with random geometries. The solutions are validated by imaging and manipulation experiments with wet and dry paper fibers and dry paper fiber bonds. In the imaging experiments, the results are compared with the reference data obtained either from an experienced human or another imaging device. The results show that these solutions provide morphological data about the fibers which is accurate and precise enough to enable automated fiber manipulation. Although this thesis is focused on the manipulation of paper fibers and paper fiber bonds, both the illumination solution and the computer vision algorithms are applicable to other types of fibrous materials

Transfer printing based microassembly and colloidal quantum dot film integration

Micro / nanoscale manufacturing requires unique approaches to accommodate the immensely different characteristics of the miniscule objects due to their high surface area to volume ratio when compared with macroscale objects. Therefore, surface forces are much more dominating than body forces, which causes the significant difficulty of miniscule object manipulation. Because of this challenge, monolithic microfabrication relying on photolithography has been the primary method to manufacture micro / nanoscale structures and devices in place of microassembly. However, by virtue of the two-dimensional (2D) nature of photolithography, formation of complex 3D shape architectures via monolithic microfabrication is inherently limited, which would otherwise enable improvements in performance and novel functionalities of devices. Furthermore, monolithic microfabrication is compatible only with materials which survive in a wet condition during photolithography. Delicate nanomaterials such as colloidal quantum dots cannot be processed via monolithic microfabrication. In this context, transfer printing has emerged as a method to transfer heterogeneous material pieces from their mother substrates to a foreign substrate utilizing a polymeric stamp in a dry condition. In this thesis, advanced modes of transfer printing are studied and optimized to enable a 3D microassembly called ‘micro-Lego’ and a novel strategy of quantum dot film integration. Micro-Lego involves transfer printing for material piece pick-and-place and thermal joining for irreversible permanent bonding of placed material pieces. A microtip elastomeric stamp is designed to advance transfer printing and thermal joining processes are optimized to ensure subsequent material bonding. The mechanical joining strength between material pieces assembled by micro-Lego are characterized by means of blister tests and the nanoindentation. Moreover, the electrical contact between two conducting materials formed by micro-Lego are examined. Lastly, inspired from the subtractive transfer printing technique, protocols of quantum dot film patterning using polymeric stamps made of a shape memory polymer as well as a photoresist are established for the convenient integration of quantum dots in various geometries and configurations as desired. Transfer printing-based micro / nanoscale manufacturing presented in this thesis opens up new pathways to manufacture not only complex 3D functional micro devices but also high resolution nano devices for unparalleled performance or for an unusual functionality, which are unattainable through monolithic microfabrication

Affordable flexible hybrid manipulator for miniaturised product assembly

Miniaturised assembly systems are capable of assembling parts of a few millimetres in size with an accuracy of a few micrometres. Reducing the size and the cost of such a system while increasing its flexibility and accuracy is a challenging issue. The introduction of hybrid manipulation, also called coarse/fine manipulation, within an assembly system is the solution investigated in this thesis. A micro-motion stage (MMS) is designed to be used as the fine positioning mechanism of the hybrid assembly system. MMSs often integrate compliant micro-motion stages (CMMSs) to achieve higher performances than the conventional MMSs. CMMSs are mechanisms that transmit an output force and displacement through the deformation of their structure. Although widely studied, the design and modelling techniques of these mechanisms still need to be improved and simplified. Firstly, the linear modelling of CMMSs is evaluated and two polymer prototypes are fabricated and characterised. It is found that polymer based designs have a low fabrication cost but not suitable for construction of a micro-assembly system. A simplified nonlinear model is then derived and integrated within an analytical model, allowing for the full characterisation of the CMMS in terms of stiffness and range of motion. An aluminium CMMS is fabricated based on the optimisation results from the analytical model and is integrated within an MMS. The MMS is controlled using dual-range positioning to achieve a low-cost positioning accuracy better than 2µm within a workspace of 4.4×4.4mm2. Finally, a hybrid manipulator is designed to assemble mobile-phone cameras and sensors automatically. A conventional robot manipulator is used to pick and place the parts in coarse mode while the aluminium CMMS based MMS is used for fine alignment of the parts. A high-resolution vision system is used to locate the parts on the substrate and to measure the relative position of the manipulator above MMS using a calibration grid with square patterns. The overall placement accuracy of the assembly system is ±24µm at 3σ and can reach 2µm, for a total cost of less than £50k, thus demonstrating the suitability of hybrid manipulation for desktop-size miniaturised assembly systems. The precision of the existing system could be significantly improved by making the manipulator stiffer (i.e. preloaded bearings…) and adjustable to compensate for misalignment. Further improvement could also be made on the calibration of the vision system. The system could be either scaled up or down using the same architecture while adapting the controllers to the scale.Engineering and Physical Sciences Research Council (EPSRC

Micro/Nano Structures and Systems

Micro/Nano Structures and Systems: Analysis, Design, Manufacturing, and Reliability is a comprehensive guide that explores the various aspects of micro- and nanostructures and systems. From analysis and design to manufacturing and reliability, this reprint provides a thorough understanding of the latest methods and techniques used in the field. With an emphasis on modern computational and analytical methods and their integration with experimental techniques, this reprint is an invaluable resource for researchers and engineers working in the field of micro- and nanosystems, including micromachines, additive manufacturing at the microscale, micro/nano-electromechanical systems, and more. Written by leading experts in the field, this reprint offers a complete understanding of the physical and mechanical behavior of micro- and nanostructures, making it an essential reference for professionals in this field