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Mechanical manipulation and characterization of biological cells have wide applications in genetics, reproductive biology, and cell mechanics. This research focuses on (1) the development of enabling microrobotic systems and techniques for automated cell microinjection and in situ mechanical characterization; and (2) the demonstration of molecule efficacy testing and cell quality assessment with the new technologies. Targeting high-speed cell injection for molecule screening, a first-of-its-kind automated microrobotic cell injection system is developed for injecting foreign materials (e.g., DNA, morpholinos, and proteins) into zebrafish embryos (& sim;1.2 mm) and mouse oocytes/embryos (& sim;100 mum), which overcomes the problems inherent in manual operation, such as long learning curves, human fatigue, and large variations in success rates due to poor reproducibility. Novel cell holding devices are developed for immobilizing a large number of embryos into a regular pattern, greatly facilitating sample preparation and increasing the sample preparation speed. Leveraging motion control and computer vision techniques, the microrobotic system is capable of performing robust cell injection at a high speed with high survival, success, and phenotypic rates. The mouse embryo injection system is applied to molecule testing of recombinant mitochondrial proteins. The efficacy of an anti-apoptotic Bcl-xL (DeltaTM) protein is, for the first time, quantitatively evaluated for enhancing the development competence of mouse embryos. For cell quality assessment, this research develops a vision-based technique for real-time cellular force measurement and in situ mechanical characterization of individual cells during microinjection. A microfabricated elastic device and a sub-pixel computer vision tracking algorithm together resolve cellular forces at the nanonewton level. Experimental results on young and old mouse oocytes demonstrate that the in situ obtained force-deformation data can be used for mechanically distinguishing healthy mouse oocytes from those with cellular dysfunctions. This work represents the first study that quantified the mechanical difference between young and old mouse oocytes, promising a practical way for oocyte quality assessment during microinjection.
Robotics for Cell Manipulation and Characterization provides fundamental principles underpinning robotic cell manipulation and characterization, state-of-the-art technical advances in micro/nano robotics, new discoveries of cell biology enabled by robotic systems, and their applications in clinical diagnosis and treatment. This book covers several areas, including robotics, control, computer vision, biomedical engineering and life sciences using understandable figures and tables to enhance readers’ comprehension and pinpoint challenges and opportunities for biological and biomedical research. Focuses on, and comprehensively covers, robotics for cell manipulation and characterization Highlights recent advances in cell biology and disease treatment enabled by robotic cell manipulation and characterization Provides insightful outlooks on future challenges and opportunities
The mechanical response of a cell to external forces carries information about its structure and function. Because cell manipulation should ideally be non-invasive while performing sophisticated biophysical characterization, the radiation force of optical tweezers has become highly attractive. In this thesis, we explore three types of recently-developed optical tweezers: 1) static, 2) time-sharing and 3) oscillating. Using a full three-dimensional finite element method (3DFEM), modeling of each of these regimes allows us to fit experiments and access the cell mechanical properties. Combining optical trapping with cell mechanics requires interdisciplinary efforts. A survey of the various experimental approaches for optical trapping and measurements on isolated cells is presented. We then lay the theoretical background linking the interaction of optical fields to the cell's mechanical response. We are the first to implement a 3DFEM calculation including light scattering and the radiation stress distribution to predict the deformation of a biconcave cell -emulating a red blood cell- in static dual-trap optical tweezers. At equilibrium, the final deformation is given by the separation distance of the two trapping beams, revealing how the cell can be elongated or shrunk. Time-sharing optical tweezers realize multiple traps to manipulate objects ranging from macromolecules to biological cells. Our quantitative analysis shows how, although jumping, the local stress and strain is omnipresent in the cell. The viscoelastic object deformation and internal energy dissipation are analyzed. Another cell shape, a cubic rod, is also studied, elucidating novel symmetrical properties of the mechanical response. Finally, the analysis of the time-dependent deformation -creep testing- of a cell in static and time-sharing optical tweezers, shows that deformation of the object depends altogether on the object's viscoelasticity, significantly on its 3D shape and the mechanical loading. However, dynamic testing with oscillating optical tweezers surprisingly shows a phase shift between the loading stress (external force) and strain (deformation) independent on the 3D cell shape. This is a novel avenue giving access to the cell's viscoelasticity dynamic complex modulus directly in the time-domain.
This book is a printed edition of the Special Issue "Single Cell Analysis in Biotechnology and Systems Biology" that was published in IJMS
This book provides an overview of single-cell isolation, separation, injection, lysis and dynamics analysis as well as a study of their heterogeneity using different miniaturized devices. As an important part of single-cell analysis, different techniques including electroporation, microinjection, optical trapping, optoporation, rapid electrokinetic patterning and optoelectronic tweezers are described in detail. It presents different fluidic systems (e.g. continuous micro/nano-fluidic devices, microfluidic cytometry) and their integration with sensor technology, optical and hydrodynamic stretchers etc., and demonstrates the applications of single-cell analysis in systems biology, proteomics, genomics, epigenomics, cancer transcriptomics, metabolomics, biomedicine and drug delivery systems. It also discusses the future challenges for single-cell analysis, including the advantages and limitations. This book is enjoyable reading material while at the same time providing essential information to scientists in academia and professionals in industry working on different aspects of single-cell analysis. Dr. Fan-Gang Tseng is a Distinguished Professor of Engineering and System Science at the National Tsing Hua University, Taiwan. Dr. Tuhin Subhra Santra is a Research Associate at the California Nano Systems Institute, University of California at Los Angeles, USA.
This book provides an overview of the noteworthy developments in the field of micromachining, with a specific focus on microinjection systems used for biological micromanipulation. The author also explores the design, development, and fabrication of new mechanical designs for micromachines, with plenty of examples that elucidate their modeling and control. The design and fabrication of a piezoelectric microinjector, constant force microinjector, constant force microgripper, PDVF microforce sensor, and a piezoelectric microsyringe are presented as examples of new technology for microinjection systems. This book is appropriate for both researchers and advanced students in bioengineering.
This volume brings together a broad array of scientific expertise to focus on the characterization and utilization of cellulosic materials. Researchers from Austria, Germany, Sweden, Japan, New Zealand, Australia, and the U.S. explore many facets of the plant cell wall, from its fundamental structure and its manipulation via molecular biology to its application in composite materials. Exciting applications of near infrared spectroscopy, x-ray diffraction, confocal microscopy, and molecular coupling as a viscoelastic probe provide new insights into the ultrastructure and properties of cellulosic materials.
Robotic Cell Manipulation introduces up-to-date research to realize this new theme of medical robotics. The book is organized in three levels: operation tools (e.g., optical tweezers, microneedles, dielectrophoresis, electromagnetic devices, and microfluidic chips), manipulation types (e.g., microinjection, transportation, rotation fusion, adhesion, separation, etc.), and potential medical applications (e.g., micro-surgery, biopsy, gene editing, cancer treatment, cell-cell interactions, etc.). The technology involves different fields such as robotics, automation, imaging, microfluidics, mechanics, materials, biology and medical sciences. The book provides systematic knowledge on the subject, covering a wide range of basic concepts, theories, methodology, experiments, case studies and potential medical applications. It will enable readers to promptly conduct a systematic review of research and become an essential reference for many new and experienced researchers entering this unique field. Introduces the applications of robot-assisted manipulation tools in various cell manipulation tasks Defines many essential concepts in association with the robotic cell manipulation field, including manipulation strategy and manipulation types Introduces basic concepts and knowledge on various manipulation devices and tasks Describes some cutting-edge cell manipulation technologies and case studies