No full-text available
To read the full-text of this research,
you can request a copy directly from the authors.
Development of a Micromanipulation Platform with Passive-Active Hybrid Release Strategy for Single-Cell Separation
... Some of these fluids, such as the tris-buffered saline used for DNA stretching [117], have relatively high salt concentrations, leading to a shortening of the Debye length and thus a decreasing importance of any stabilising electrostatic interactions [118]. Methods for reducing adhesion forces in microrobotics are based around reducing contact area between objects [112,119]. The reason for this can be easily appreciated when one considers the equations for the van der Waals force between a sphere and a plane (11) [120]-a key contributor to adhesion forces. ...
Optical tweezers have been used for biological studies since shortly after their inception. However, over the years research has suggested that the intense laser light used to create optical traps may damage the specimens being studied. This review aims to provide a brief overview of optical tweezers and the possible mechanisms for damage, and more importantly examines the role of optical micromachines as tools for biological studies. This review covers the achievements to date in the field of optical micromachines: improvements in the ability to produce micromachines, including multi-body microrobots; and design considerations for both optical microrobots and the optical trapping set-up used for controlling them are all discussed. The review focuses especially on the role of micromachines in biological research, and explores some of the potential that the technology has in this area.
Cells play significant roles in our day to day life. However the interactions of cells to cells and responses of organelles to molecules and intracellular behaviours are still not fully understood. To understand better physiological interactions among molecules, organelles, and cells, ensemble average measurement for millions of cells together is not detailed enough to provide the information in single cell level. For example, the biological function such as genome, epigenome, and transcriptome at bulk population may be informative, however it is not enough to provide the cell heterogeneity characteristics in phenotypic behaviors and molecular dynamics. Again it cannot provide any information of an underrepresented cell subpopulation that could have a differential or crucial function in a specific biological context, such as stem cells or tumor initiation cells. In contrast, single cell sequencing (SCS) is able to empirically infer the driver mutations and map the sequential mutation events during cancer development. The integration of genomics and transcriptomics in single cancer cells will also provide valuable information on the functional consequences of mutations and copy number variations in these cells. Isolated single cells can improve whole genome, transcriptome amplification and genome-wide analysis platform through advanced sequencing techniques, which not only allow high resolution genome and transcriptome analysis, but also it have potential to reveal the epigenome map of the target single cells. Undoubtedly, these novel approaches will produce profound health benefits, such as a more efficient treatment strategy for genetic disorders patients, which can be realized by revealing at single cell level.
Apart from the powerful possibility for single cell analysis (SCA), the huge data generated from SCA process has also been emerging as a challenging issue. In recent years, bioinformatics technique has been employed to study “big data” from large ensembles of single cell data. Thus interestingly, the unresolved questions in the past, such as whether any of two single cells are really the same, if we are able to measure the parameters with sufficient accuracy? Is there two cells have similar biological function with predictable outcomes, if we treat cells with same drugs or environmental factors? it may be partially answered now, thanks to the unique information obtained from Single cell analysis. Thus, SCA, without doubt, is an efficient and valuable approach to understand the fundamental biology in embryonic development, detailed cell lineage tree in higher organisms, or dissection of tumor heterogeneity and disease, etc.
To analyse the cellular function, SCA can be performed by combining capillary electrophoresis (CE) with laser induced fluorescence (LIF) detection, electrochemical detection (ED), flow cytometry or mass spectrometry etc. Recently the development of MEMS (Micro Electro Mechanical Systems) technology with integration of chemical engineering, chemistry, and life science with micro/nanofluidic devices to become Bio-MEMS, Lab on a Chip, or micro total analysis systems (µTAS), which can enable more complex manipulations of chemicals and biological agents in micro/nano fluidic environments. Micro/nanofluidic devices with the power to manipulate and detect bio-samples, reagents, or biomolecules at micro/nano scale can well fulfill the requirements for single cells analysis. Thus, micro-nanofluidic devices not only useful for cell manipulation, cell isolation, cell lysis, cell separation, but also it can easily control the biochemical, electrical, mechanical parameters for SCA analysis with a precise control of the dosage inside single cell, spatial resolution, or temporal pace.
This book provides an overview of single cell isolation, injection, lysis and dynamic analysis as well as their heterogeneity study by using different miniaturized devices. As an important part SCA, different methods including electroporation, microinjection, optoporation or photoporation are introduced in detail. Different fluidic systems (e.g. continuous micro/nano-fluidic devices, microfluidic cytometry) and their integration with sensor technology, optical and hydrodynamic stretcher etc. are also covered. The role of single cell analysis in system biology, genomics, epigenomics, proteomics, cancer transcriptomics, metabolomics, the applications of single cell analysis for bio- catalysis, metabolic as well as bioprocess engineering and the future challenges of single cell analysis with their advantage as well as their limitations are also discussed. The book contain fifteen chapters and each chapter provides a short abstract, a brief introduction, related experimental methods, detail analysis schemes, conclusions, and some comments on the technology advancement/limitation for single cell analysis. Chapter 1 describes single cell behavioral assay for heterogeneity study by single cell isolation and tracking to investigate cell proliferation, differentiation and lineage. There are two platforms discussed, including cell migration platforms to measure cell motility, deformation, invasiveness, and cell-cell interaction platforms to study the alteration of cell behaviors caused by reciprocal interactions among cells. Chapter 2 presents single cell analysis in system biology, which enable the isolation of individual cells in a form that accommodates systomics studies of the biological methods and then deployed on such isolated cells to generate system-level information. Finally it describes the bioinformatics technique that is specifically directed towards single-cell studies. Chapter 3 presents advanced single cell electroporation and localized single cell electroporation technique with intracellular delivery and lysis by using different geometry based micro/nanofluidic devices. Chapter 4 focuses on single cell microinjection technique which intended to provide a basic knowledge of the microinjection, advantages, disadvantages and its development, applications, basic instrumentation required along with a basic protocol and its uses. Chapter 5 discuses optical tools for single cell analysis ranging from optical trapping systems which provide a contact-free technique for the manipulation of micron-scale objects to a selection of different optically-mediated cell membrane disruption methods available for lysis and/or delivery of material. Two newly-developed optoelectrokinetic techniques, termed rapid electrokinetic patterning (REP) and optoelectronic tweezers (OETs) with their detail setup, fabrication, assessment and the fundamental principles to their applications in cell-related research are discussed in Chapter 6. Chapter 7 describe continuous micro/nano-fluidic devices for single cell analysis which broadly divided into two parts—single-cell manipulation (SCM) and single-cell analysis (SCA). The first part of the chapter presents state-of-the-art techniques developed to handle single cells, including counting, sorting, positioning, and culturing, which are essential steps in many biological and medical assays. The second part of this chapter describe manipulation combine with other stimulating and sensing techniques for the observation and characterization of single cells. Chapter 8 emphasize the relationships between cellular mechanical properties and various disease processes and changes in cell states. The mechanical properties of single cells is measured using atomic force microscopy and micropipette aspiration. Finally the microfluidic approaches including microfluidic constriction channels, microfluidic optical stretchers and microfluidic hydrodynamic stretchers, which are being developed as next-generation, automated, and high-throughput techniques for characterization of the mechanical properties of single cells. The advantages and limitations of each technique are compared and future research opportunities also highlighted. Chapter 9 presents cytometry of single cells for biology and biomedicine where it describe the advantages of combining different approaches in some kind of integrated instrument that could perform both flow cytometry and image analysis on single cells as well as examining the internal contents of each single cell. Chapter 10 discusses the role of genomics and epigenomics in single cell where it briefly describe the major technological developments achieved in single cell “omics” as well as the technical challenges to overcome and the potential of future developments. This chapter also discuss the breakthroughs of single cell “omics” with a special focus on the integration of all biological methods to understand whole-organism biology. Chapter 11 focuses on single cell metabolomics in system biology, where the chapter emphasize the recent improvement of analytical tools to unravel single cell metabolomics, as well as their specificity. This includes the exciting development and expansion of analytical tool technologies in metabolite analysis. Chapter 12 presents different types of cell based drug delivery systems to facilitate treatments for infectious and noninfectious diseases, potentiality and limitations of single cell assay along with the clinical aspects of cell based drug delivery. Chapter 13 describes single cell sequencing (SCS) methodologies, existing applications especially its capability in reconstructing the evolutionary history of cancer progression and in profiling cancer transcriptome. Chapter 14 discusses single cell characterization of Microalgal lipid contents with confocal Raman microscopy resulted in remarkable enhancements in the sensitivity, specificity, and spatio-temporal resolution for analyzing lipid content of algal isolates obtained through a mutagenesis screen of the green alga, Chlamydomonas reimhardtii, for increased lipid production at the single cell level. In last Chapter 15 demonstrates single-cell DNA devices to improve DNA delivery accuracy for retinoic acid-induced P19 neurons under optimal conditions with a mathematical modeling and physical hypothesis. Also the chapter describe a motion model based on parameters from dynamic transport, including an anterograde state, a retrograde state, and a pausing state.
This book includes 15 chapters, and covers a wide spectrum of the essential aspects of
single cell analysis. All of these chapters are written by scientific experts in their own fields and provide technical tips based on valuable experience and knowledge. Potential problems and challenges as well as possible solutions are also provided with an emphasis on the future prospectus. We hope this book can be an enjoyable reading material and abundant resource to provide essential information to those scientists in academia and engineers in industry working on different aspects of SCA. .
Editor
Dr. Fan-Gang Tseng
Dr. Tuhin Subhra Santra
Due to scale effects, microoperation, especially the releasing of microobjects, has been a long-standing challenge in micromanipulation applications. In this paper a micromanipulation method is presented based on dynamic adhesion control with compound vibration. This adhesion control technique employs inertia force to overcome adhesion force achieving 100% repeatability with releasing accuracy of 4 +/- 0.5 mu m, which was experimentally quantified through the manipulation of 20-100 mu m polystyrene spheres under an optical microscope. The micromanipulation system consists of a microgripper and a piezoelectric ceramics module. The compound vibration comes from the electrostatic actuator and the piezoelectrically driven actuator. Surface and bulk micromachining technology is employed to fabricate the microgripper used in the system from a single crystal silicon wafer. Experimental results confirmed that this adhesion control technique is independent of substrate. Theoretical analyses were conducted to understand the picking up and releasing mechanism. Based on this preliminary study, the micromanipulation system proved to be an effective solution for active picking up and releasing of micromanipulation.
As mechanical end-effectors, microgrippers enable the pick–transport–place of micrometer-sized objects, such as manipulation and positioning of biological cells in an aqueous environment. This paper reports on a monolithic MEMS-based microgripper with integrated force feedback along two axes and presents the first demonstration of force-controlled micro-grasping at the nanonewton force level. The system manipulates highly deformable biomaterials (porcine interstitial cells) in an aqueous environment using a microgripper that integrates a V-beam electrothermal microactuator and two capacitive force sensors, one for contact detection (force resolution: 38.5 nN) and the other for gripping force measurements (force resolution: 19.9 nN). The MEMS-based microgripper and the force control system experimentally demonstrate the capability of rapid contact detection and reliable force-controlled micrograsping to accommodate variations in size and mechanical properties of objects with a high reproducibility.
The atomic force microscope (AFM) has been successfully used to perform nanorobotic manipulation operations on nanoscale entities such as particles, nanotubes, nanowires, nanocrystals, and DNA since 1990s.
There have been many progress on modeling, imaging, teleoperated or automated control, human-machine interfacing, instrumentation, and applications of AFM based nanorobotic manipulation systems in literature. This book aims to include all of such state-of-the-art progress in an organized, structured, and detailed manner as a reference book and also potentially a textbook in nanorobotics and any other nanoscale dynamics, systems and controls related research and education.
Clearly written and well-organized, this text introduces designs and prototypes of the nanorobotic systems in detail with innovative principles of three-dimensional manipulation force microscopy and parallel imaging/manipulation force microscopy.
Object handling has been a challenging operation at the microscale level. In order to manipulate these micro-objects within a confined workspace, it is inevitably imperative for the handling tool to be manipulated along an unobstructed trajectory or path. In addition, the presence of undesirable adhesion force can cause the object to adhere to the tool, making the release process problematic. This work presents the development of an automated dual-arm micromanipulation process to overcome these challenges in micro-object handling. The two probes of the dual-arm micromanipulation system were individually controlled to manipulate a micro-sphere lying on top of a glass substrate. A potential field path-planning algorithm was employed to evaluate the paths for positioning of the two probes and prevent the probes from undergoing collisions. To release the grasped micro-sphere after manipulation with minimal adhesion force, the orientations of two probes were reconfigured to reduce the contact area with the sphere. Vision images obtained from a top-view CCD camera were utilized to evaluate the contour of the sphere for automatic reconfiguration of the two probes. Experimental results confirmed that the two probes can be manipulated to grasp the targeted micro-sphere from arbitrary positions and the incorporation of the probe reconfiguration scheme yields a higher success rate for sphere release.
In this paper, a dynamic releasing approach is proposed for high-speed biological cell manipulation. A compact parallel mechanism for grasping and releasing microobjects is used to generate controllable vibration to overcome the strong adhesion forces between the end effector and the manipulated object. To reach the required acceleration of the end effector, which is necessary for the detachment of the target object by overcoming adhesion forces, vibration in the end effector is generated by applying sinusoidal voltage to the PZT actuator of the parallel mechanism. For the necessary acceleration, we focus on the possible range of the frequency of the PZT-actuator-induced vibration, while minimizing the amplitude of the vibration (14 nm) to achieve precise positioning. The effect of the air and liquid environments on the required vibration frequency for successful release is investigated. For the first time, release results of microbeads and biological cells are compared. Release of the biological cells with 100 % success rate suggests that the proposed active release method is an appropriate solution for adhered biological cells during the release task.
Micro- and nanosized objects aligned in specific spatial order are of great interest for applications in photonics and nanoelectronics. In particular, piezo-actuated robotic setups are promising tools to arrange and manipulate these objects individually. However, automated robotic processing on the submicron
scale remains challenging due to the force scaling laws and the limited possibilities in terms of control. This paper presents the current progress on fully-automated pick-andplace routines of individual colloidal particles using a dedicated dual-probe setup inside a scanning electron microscope. Applying tailored probes in combination with image processing of the visual feedback provided by the microscope allow for complex
automation sequences. The limits of the current technique are highlighted and the challenges for automated processing of progressively smaller particles are discussed.
An 8-DOF micro hand consisting of two rotational fingers is developed. Each finger has 4 DOF: 3-DOF translation of the fingertip driven by three linear stages and 1-DOF rotation about the finger’s cylindrical axis driven by one rotation stage. The finger is a glass rod attached to the rotation stage. The two fingers are tilted and placed on an inverted microscope’s stage from both sides of the objective lens. The fingertips cross each other and grasp an object above the objective lens. The fingers translate the grasped object spatially by moving in the same direction together. The fingers can also rotate the object about its two axes if the fingers move along their cylindrical axes in the opposite directions or rotate about their cylindrical axes. Because of the eccentricity of rotation, the finger cannot rotate exactly on its cylindrical axis only using the rotation stage. This error is cancelled by moving the three linear stages in the opposite directions to the error. Virtual stick method for manual operation of this micro hand is implemented. A human operator commands 3-DOF translation and 2-DOF rotation of the grasped object using a PHANTOM omni-device, as if the operator directly manipulates the stick which is virtually fixed to the center of the object. The commanded motion of the object is converted to the motions of the two fingers, which are inputs to the controllers of the fingers. The developed micro hand can grasp and translate a spherical soft object of about 120[μm] in diameter spatially by virtual stick method. The hand can also rotate the object about its two axes by virtual stick method.
This paper gives a review of existing models used by other authors aiming at modeling micromanipulation tasks. It introduces the distinction between surface forces, which act even at distance (van der Waals (VDW), capillary and electrostatic forces) and contact forces, which are closely related to deformation and adhesion. Moreover, it presents our work on VDW and capillary forces: compared to existing approximations, these models allow to take more parameters into account such as, for example, statistical roughness in VDW forces or the volume of liquid in capillary forces. They could be used, for example, to build up new handling strategies as illustrated in the references cited in the paper. However, this paper focuses on fundamental models and does not present any specific microhandling strategy.
Due to force scaling laws, large adhesion forces at the microscale make rapid accurate release of microobjects a long-standing challenge in pick-place micromanipulation. This paper presents a new microelectromechanical systems (MEMS) microgripper integrated with a plunging mechanism to impact the microobject for it to gain sufficient momentum to overcome adhesion forces. The performance was experimentally quantified through the manipulation of 7.5-10.9-mum borosilicate glass spheres in an ambient environment under an optical microscope. Experimental results demonstrate that this microgripper, for the first time, achieves a 100% successful release rate (based on 200 trials) and a release accuracy of 0.70 plusmn0.46 mum. Experiments with conductive and nonconductive substrates also confirmed that the release process is not substrate dependent. Theoretical analyses were conducted to understand the release principle. Based on this paper, further scaling down the end structure of this microgripper will possibly provide an effective solution to the manipulation of submicrometer-sized objects.
During microscale object manipulation, contact (pull-off) forces and non-contact (capillary, van der Waals and electrostatic) forces determine the behaviour of the micro-objects rather than the inertial forces. The aim of this article is to give an experimental analysis of the physical phenomena at a microscopic scale in dry and liquid media. This article introduces a review of the major differences between dry and submerged micromanipulations. The theoretical influences of the medium on van der Waals forces, electrostatic forces, pull-off forces and hydrodynamic forces are presented. Experimental force measurements based on an AFM system are carried out. These experiments exhibit a correlation better than 40 % between the theoretical forces and the measured forces (except for pull-off in water). Finally, some comparative experimental micromanipulation results are described and show the advantages of the liquid medium.
This paper presents a computer vision algorithm for visually detecting the contact between an end-effector and a target surface under an optical microscope. Without using additional sensors (e.g., proximity or force/touch sensors), this algorithm provides robustness and a sub-micrometer detection resolution. Fundamentally, after the establishment of a contact in the world frame, further vertical motion induces horizontal motion in the image plane. An analysis is presented to elaborate this fundamental mechanism. Experimental results demonstrate that this computer-vision-based method is capable of achieving contact detection between a micropipette and a glass slide surface with a resolution of 0.2 microm. Furthermore, 1300 experimental trials reveal that the presented algorithm is robust to variations in illumination intensity, microscopy magnification, and microrobot motion speed.
When parts to be handled are less than one millimeter in size, adhesive forces between gripper and object can be significant compared to gravitational forces. These adhesive forces arise primarily from surface tension, Van der Waals, and electrostatic attractions and can be a fundamental limitation to part handling in a gas environment. While it is possible to fabricate miniature versions of conventional robot grippers, for example from polysilicon, it appears that it will be difficult to overcome adhesion effects for the smallest parts. Thus, manipulation of parts on the order of 10 micron or smaller may best be done in a fluid medium using techniques such as laser trapping, or dielectrophoresis