A Fully Automated Robotic System for Microinjection of Zebrafish Embryos

As an important embodiment of biomanipulation, injection of foreign materials (e.g., DNA, RNAi, sperm, protein, and drug compounds) into individual cells has significant implications in genetics, transgenics, assisted reproduction, and drug discovery. This paper presents a microrobotic system for fully automated zebrafish embryo injection, which overcomes the problems inherent in manual operation, such as human fatigue and large variations in success rates due to poor reproducibility. Based on computer vision and motion control, the microrobotic system performs injection at a speed of 15 zebrafish embryos (chorion unremoved) per minute, with a survival rate of 98% (n = 350 embryos), a success rate of 99% (n = 350 embryos), and a phenotypic rate of 98.5% (n = 210 embryos). The sample immobilization technique and microrobotic control method are applicable to other biological injection applications such as the injection of mouse oocytes/embryos and Drosophila embryos to enable high-throughput biological and pharmaceutical research

Haptic guidance for microrobotic intracellular injection

The ability for a bio-operator to utilise a haptic device to manipulate a microrobot for intracellular injection offers immense benefits. One significant benefit is for the bio-operator to receive haptic guidance while performing the injection process. In order to address this, this paper investigates the use of haptic virtual fixtures for cell injection and proposes a novel force field virtual fixture. The guidance force felt by the bio-operator is determined by force field analysis within the virtual fixture. The proposed force field virtual fixture assists the bio-operator when performing intracellular injection by limiting the micropipette tip\u27s motion to a conical volume as well as recommending the desired path for optimal injection. A virtual fixture plane is also introduced to prevent the bio-operator from moving the micropipette tip beyond the deposition target inside the cell. Simulation results demonstrate the operation of the guidance system.<br /

3D particle-based cell modelling for haptic microrobotic cell injection

Introducing haptic interface to conduct microrobotic intracellular injection has many beneficial implications. In particular, the haptic device provides force feedback to the bio-operator\u27s hand. This paper introduces a 3D particle-based model to simulate the deformation of the cell membrane and corresponding cellular forces during microrobotic cell injection. The model is based on the kinematic and dynamic of spring &ndash; damper multi particle joints considering visco-elastic fluidic properties. It simulates the indentation force feedback as well as cell visual deformation during the microinjection. The model is verified using experimental data of zebrafish embryo microinjection. The results demonstrate that the developed cell model is capable of estimating zebrafish embryo deformation and force feedback accurately

Novel parameter estimation schemes in microsystems

This paper presents two novel estimation methods that are designed to enhance our ability of observing, positioning, and physically transforming the objects and/or biological structures in micromanipulation tasks. In order to effectively monitor and position the microobjects, an online calibration method with submicron precision via a recursive least square solution is presented. To provide the adequate information to manipulate the biological structures without damaging the cell or tissue during an injection, a nonlinear spring-mass-damper model is introduced and mechanical properties of a zebrafish embryo are obtained. These two methods are validated on a microassembly workstation and the results are evaluated quantitatively

Three dimensional mechanical modelling and analysis of embryos microinjection.

This research has developed a novel 3D particle based cell model that is able to accurately estimate the membrane deformation and indentation force in bio-micromanipulation of an individual cell, especially embryos microinjection practice

Microdevices and Microsystems for Cell Manipulation

Microfabricated devices and systems capable of micromanipulation are well-suited for the manipulation of cells. These technologies are capable of a variety of functions, including cell trapping, cell sorting, cell culturing, and cell surgery, often at single-cell or sub-cellular resolution. These functionalities are achieved through a variety of mechanisms, including mechanical, electrical, magnetic, optical, and thermal forces. The operations that these microdevices and microsystems enable are relevant to many areas of biomedical research, including tissue engineering, cellular therapeutics, drug discovery, and diagnostics. This Special Issue will highlight recent advances in the field of cellular manipulation. Technologies capable of parallel single-cell manipulation are of special interest

Novel estimation and control techniques in micromanipulation using vision and force feedback

With the recent advances in the fields of micro and nanotechnology, there has been growing interest for complex micromanipulation and microassembly strategies. Despite the fact that many commercially available micro devices such as the key components in automobile airbags, ink-jet printers and projection display systems are currently produced in a batch technique with little assembly, many other products such as read/write heads for hard disks and fiber optics assemblies require flexible precision assemblies. Furthermore, many biological micromanipulations such as invitro-fertilization, cell characterization and treatment rely on the ability of human operators. Requirement of high-precision, repeatable and financially viable operations in these tasks has given rise to the elimination of direct human involvement, and autonomy in micromanipulation and microassembly. In this thesis, a fully automated dexterous micromanipulation strategy based on vision and force feedback is developed. More specifically, a robust vision based control architecture is proposed and implemented to compensate errors due to the uncertainties about the position, behavior and shape of the microobjects to be manipulated. Moreover, novel estimators are designed to identify the system and to characterize the mechanical properties of the biological structures through a synthesis of concepts from the computer vision, estimation and control theory. Estimated mechanical parameters are utilized to reconstruct the imposed force on a biomembrane and to provide the adequate information to control the position, velocity and acceleration of the probe without damaging the cell/tissue during an injection task

Haptic technology for micro-robotic cell injection training systems — a review

Currently, the micro-robotic cell injection procedure is performed manually by expert human bio-operators. In order to be proficient at the task, lengthy and expensive dedicated training is required. As such, effective specialized training systems for this procedure can prove highly beneficial. This paper presents a comprehensive review of haptic technology relevant to cell injection training and discusses the feasibility of developing such training systems, providing researchers with an inclusive resource enabling the application of the presented approaches, or extension and advancement of the work. A brief explanation of cell injection and the challenges associated with the procedure are first presented. Important skills, such as accuracy, trajectory, speed and applied force, which need to be mastered by the bio-operator in order to achieve successful injection, are then discussed. Then an overview of various types of haptic feedback, devices and approaches is presented. This is followed by discussion on the approaches to cell modeling. Discussion of the application of haptics to skills training across various fields and haptically-enabled virtual training systems evaluation are then presented. Finally, given the findings of the review, this paper concludes that a haptically-enabled virtual cell injection training system is feasible and recommendations are made to developers of such systems