Extraction of Depth Information and Image Processing in Manipulator Cell Injection

在细胞注射时,显微镜上CCD获取的只是注射针的二维信息,但光轴方向的深度信息丢失,这会导致细胞注射的失败,必须用其他方法获得。利用激光三角测量法对注射针以及细胞载玻片的深度信息进行测量,通过纳米平台的移动来标定激光入射角参数,对获取的数据进行图形化处理,确定线激光条纹图像间的偏移距离,从而获得标准物体高度与激光条纹图像偏移量的像素比值,该比值可以用来计算实际物体的高度。另外,利用三自由度机械手,对测量的探针离细胞载玻片的距离进行验证,获得激光三角测量法的误差值。实验结果表明,在细胞注射中,利用激光三角测量法获得注射针的深度信息是可行的。Only two dimensions information of injection needle could be captured by CCD in microscope image,and the lack of the depth information through the optical axis of microscope system would result in the failure of cell injection.A new way called laser triangulation to exract depth information of the injection needle and the slide glass was presented.The measuring principle was calibrating the projection angle of laser,θ was callibrated by the remotion of nano platform,then ascertain the displacement of laser stripe,S by image processing,The height of actual objects will be decided by S· tgθ.In addition,the error of laser triangulation is demonstrated to be about from 5 to 10 μm by calibration using a 3-DOF manipulators.The result indicates that it is feasible to measure the depth information of injection needle by laser triangulation with the required precision in cell injection

Fast identification algorithms for manipulating biological cells

The physical manipulation of biological cells is very attractive now in biotechnology (Butler, 1991)) because it opens the possibility of examining and manipulating single molecules. Other methods are based on chemical effects, electrical effects, etc., and they generally do not allow researchers to examine single molecules cell and, thus, to understand their interaction which may encode many useful pieces of information. Such physical manipulation is fully performed by robotic devices. In order to automate the process of physical manipulation, micro machine vision for the fast identification of the objects involved is required. Typical objects that are involved are cells, cell elements, holders and injectors. In the research described in this thesis, which was carried out in the Advanced Engineering Design Laboratory of the Mechanical Engineering Department, University of Saskatchewan, algorithms for the three objects (the cell, holder and injector) were developed, implemented and tested. The results obtained have shown that the fastest identification times for these three objects are respectively 0.12s for the cell oocyte, 6.78s/100 frames for the holder, and 6.72s/100 frames for the injector. These performances are acceptable in the context of the physical manipulation of biological cells. The goal of the research described in this thesis was to develop algorithms that would give a fast recognition of the cell manipulation system. With the aid of the algorithms, an automatic operation of the cell manipulation system would be achieved. Image process and pattern recognition techniques were used in developing the Visual C++ GUI algorithms that would automatically recognize the components of the cell manipulation system for the purpose of manipulating the cells

Design and implementation of a vision system for microassembly workstation

Rapid development of micro/nano technologies and the evolvement of biotechnology have led to the research of assembling micro components into complex microsystems and manipulation of cells, genes or similar biological components. In order to develop advanced inspection/handling systems and methods for manipulation and assembly of micro products and micro components, robust micromanipulation and microassembly strategies can be implemented on a high-speed, repetitive, reliable, reconfigurable, robust and open-architecture microassembly workstation. Due to high accuracy requirements and specific mechanical and physical laws which govern the microscale world, micromanipulation and microassembly tasks require robust control strategies based on real-time sensory feedback. Vision as a passive sensor can yield high resolutions of micro objects and micro scenes along with a stereoscopic optical microscope. Visual data contains useful information for micromanipulation and microassembly tasks, and can be processed using various image processing and computer vision algorithms. In this thesis, the initial work on the design and implementation of a vision system for microassembly workstation is introduced. Both software and hardware issues are considered. Emphasis is put on the implementation of computer vision algorithms and vision based control techniques which help to build strong basis for the vision part of the microassembly workstation. The main goal of designing such a vision system is to perform automated micromanipulation and microassembly tasks for a variety of applications. Experiments with some teleoperated and semiautomated tasks, which aim to manipulate micro particles manually or automatically by microgripper or probe as manipulation tools, show quite promising results

複数の微小流を用いたマイクロマニピュレーション

電気通信大学201

Mechanical Manipulation and Characterization of Biological Cells

Mechanical manipulation and characterization of an individual biological cell is currently one of the most exciting research areas in the field of medical robotics. Single cell manipulation is an important process in intracytoplasmic sperm injection (ICSI), pro-nuclei DNA injection, gene therapy, and other biomedical areas. However, conventional cell manipulation requires long training and the success rate depends on the experience of the operator. The goal of this research is to address the drawbacks of conventional cell manipulation by using force and vision feedback for cell manipulation tasks. We hypothesize that force feedback plays an important role in cell manipulation and possibly helps in cell characterization. This dissertation will summarize our research on: 1) the development of force and vision feedback interface for cell manipulation, 2) human subject studies to evaluate the addition of force feedback for cell injection tasks, 3) the development of haptics-enabled atomic force microscope system for cell indentation tasks, 4) appropriate analytical model for characterizing the mechanical property of mouse embryonic stem cells (mESC) and 5) several indentation studies on mESC to determine the mechanical property of undifferentiated and early differentiating (6 days under differentiation conditions) mESC. Our experimental results on zebrafish egg cells show that a system with force feedback capability when combined with vision feedback can lead to potentially higher success rates in cell injection tasks. Using this information, we performed experiments on mESC using the AFM to understand their characteristics in the undifferentiated pluripotent state as well as early differentiating state. These experiments were done on both live as well as fixed cells to understand the correlation between the two during cell indentation studies. Our results show that the mechanical property of undifferentiated mESC differs from early differentiating (6th day) mESC in both live and fixed cells. Thus, we hypothesize that mechanical characterization studies will potentially pave the way for developing a high throughput system with force feedback capability, to understand and predict the differentiation path a particular pluripotent cell will follow. This finding could also be used to develop improved methods of targeted cellular differentiation of stem cells for therapeutic and regenerative medicine

マイクロ視野における画像計測およびその応用に関する研究

宇都宮大学博士（工学）学位論文・平成24年3月22日授与（甲第353号）平成23年