在仪器科学、微纳米技术与生物工程的交叉领域内，微细操作作为关键性技术之一，被用于细胞核移植和克隆、胞内信息分析、显微授精以及辅助孵化等方面。本文将功率超声驱动和传统显微操作技术相结合，提出了一种超声微细操作的方法，可用于单细胞或微小组织的穿刺、注射和吸引等微细操作。 在超声谐振驱动方式下，通过对微探针振幅、加速度和穿刺力的数值计算和分析，从理论上证明了利用超声进行微细操作的可行性。对鱼卵细胞、人体细胞和禽类卵细胞的穿刺、吸引和注射实验，验证了超声微细操作在实际应用中的有效性。利用20000帧/秒的高速拍摄技术获得穿刺实验的瞬时图像，显示并证明了超声微细操作方法在减小细胞受压变形、降低损伤，提高成功率等方面的优势。 根据相关理论，建立了超声微细操作系统的设计方法并搭建了实验系统。采用四端网络法和有限元法优化设计并制作了三种类型的微细操作换能器，分别用于不同体积生物目标的穿刺、吸引和注射，并通过导纳、振幅等参数测试方法实现其性能表征。研究了反应电压、溶液浓度和腐蚀深度三个工艺参数对电化学方法制作微探针的影响，制作了特征尺度为数百纳米的微探针。机械式双层压力调节装置、双微操作器和高速显微摄像设备的研究和组建保证了系统的正常运转和操作状态。基于大功率运放和自适应滤波技术研制的UPC1000型超声功率信号源，具有良好的驱动能力和自动频率跟踪功能。 根据超声微细操作系统的设计需求和实验结果，本文分析了负载对微细操作换能器电特性的影响，根据力—电—声的类比关系，拓展了带负载超声换能器的等效电路模型，揭示了换能器输入电特性在负载影响下的变化规律。实验证明，该模型可预测换能器输入电特性随负载的波动情况。所提出的理论模型可用于同类换能器的工况监测、驱动信号频率的自动调节等方面。
Micro manipulation is an important technology in the fields of instrument science, micro-nano technology and bioengineering. It has been used to nuclear transplantation, clone, inner cell information analysis, intracytoplasmic sperm injection, and assisted hatching. Combining the power ultrasonic driving technology with the traditional micro-manipulation technology, a novel technology of ultrasonic precise manipulation was presented in this paper. This technology can be used to penetration, injection, and suction for single cell or small tissue.By ultrasonic resonance driving, the numerical analyses and comparisons of micro probe’s amplitude, acceleration and puncture force proved the feasibility of ultrasonic precise manipulation in theory. A series of experiments on the fish eggs, human cells and bird oocytes demonstrated the validity of ultrasonic precise manipulation in practice. The instantaneous penetration images captured by high speed photography (20000fps) implied the advantages of ultrasonic precise manipulator: reduced deformation, smaller harm, higher success rate.A set of ultrasonic precise manipulation system which included six main parts was designed and established. According to four terminal network and finite element analysis, three kinds of the precise transducers which functions were penetration, injection, and suction to different targets respectively, were designed and fabricated. The performances of the precise transducers were tested by measurements of their admittance and amplitude. The micro probe which dimension was near several hundreds nanometer was processed and shaped by electrochemistry principle. The optimum experimental parameters were analyzed and presented, including reaction voltage, solution concentration, and immersion deepness. Three kinds of important supplementary instruments, which were mechanical double layer pressure regulator, two micro manipulator, and high speed photography system, perfected the operation of system. Based on a high-power operational amplifier and self-adapting filtering technology, an UPC1000 power ultrasonic signal source which has powerful driving ability and frequency tracing function was designed and completed.Based on the principle of electrics - mechanics - acoustics analogy, an equivalent electrical model of ultrasonic transducer with external load was established. This model revealed the shift rules of inputting electrical properties of an ultrasonic transducer working in the different load conditions. The experiments demonstrated that the inputting electrical properties of ultrasonic transducer can be estimated with the changes of the operational environment. For similar ultrasonic transducer, this equivalent circuit model can be used for the estimation of work condition and self-regulation of driving signal.
