Primordial germ cells transplantation is a unique approach for conservation and reconstitution of endangered fish species. This study aimed to establish techniques to culture dechorionated embryos in different incubation systems and also to determine anaesthetic concentration for fish recipients in the larval stage for subsequent primordial germ cell transplantation. Intact and dechorionated embryos were divided into three incubation systems: (1) a control group with manual replacement of the solution; (2) a closed environment with high oxygen with manual replacement of the solution; and (3) constant solution recirculation. This combination resulted in six treatments. For the evaluation of anaesthetics for larvae, the concentrations evaluated were 19.5 mM, 24.4 mM, 29.3 mM, and 34.2 mM of 2-phenoxyethanol. Anaesthesia concentration and recovery at different stages were evaluated. For transplantation, primordial germ cells of Astyanax altiparanae were transplanted into anaesthetised larvae (1 dph) of Prochilodus lineatus. Better results were obtained in the recirculation system for dechorionated embryos of P. lineatus for hatching (54.18%) and normal morphology (50.06%). The 2-phenoxyethanol anaesthetic with a dose of 29.3 mM resulted in shorter induction times, in addition to the recovery time between 5 and 10 min. By using this anaesthetic concentration at transplantation, GFP-positive cells were seen in two recipients, but the cells did not proliferate. This study established an effective incubation system for the development of the dechorionated embryo and determined an effective anaesthetic concentration for P. lineatus larvae. In addition, micromanipulation and transplantation of primordial germ cells in neotropical species were conducted for the first time.

Electromagnetic actuation systems (EMA) have excelled themselves in microscale manipulation. Yet, the fastened structure of the current systems tethers the controlled workspace. In this paper, a new electromagnetic actuation principle is investigated. The actuator structure consists of a pair of coaxially movable electromagnets integrated to a robotic manipulator. The pair induces a coaxial homogeneous magnetic field or gradient to control the magnitude of the magnetic torque or force by changing the distance between the electromagnets asymmetrically. The robotic manipulator, on the other hand, transports the pair at five degrees of freedom to manipulate a microrobot in 3D space by closed-loop control with integrated vision feedback system. Numerical analyses are performed to investigate the induced electromagnetic field at the symmetrical/asymmetrical configuration of the coaxial pair. Accordingly, a correlation between the magnitude of the magnetic force and the asymmetric distance is obtained for flexible force control. A proof of concept prototype is constructed to validate the proposed actuation principle and evaluate its performance experimentally. The experimental results verify the numerical analysis and show the system applicability of inducing controlled forces on a micro-object in 2D and 3D workspaces at a velocity range of 65 to 157 $\mu$ m/s. Moreover, micromanipulation on a helical route is also demonstrated with an absolute error mean from the reference path of 191 $\mu$ m.

The ability to analyze nanoparticles in the atom probe has often been limited by the complexity of the sample preparation. In this work, we present a method to lift–out single nanoparticles in the scanning electron microscope. First, nanoparticles are dispersed on a lacey carbon grid, then positioned on a sharp substrate tip and coated on all sides with a metallic matrix by physical vapor deposition. Compositional and structural insights are provided for spherical gold nanoparticles and a segregation of silver and copper in silver copper oxide nanorods is shown in 3D atom maps. Using the standard atom probe reconstruction algorithm, data quality is limited by typical standard reconstruction artifacts for heterogeneous specimens (trajectory aberrations) and the choice of suitable coatings for the particles. This approach can be applied to various unsupported free-standing nanoparticles, enables preselection of particles via correlative techniques, and reliably produces well-defined structured samples. The only prerequisite is that the nanoparticles must be large enough to be manipulated, which was done for sizes down to ~50 nm.

This paper presents and compares a set of calibration strategies useful to calibrate vision-based robotised work-cells for micromanipulation and microassembly. To grasp and release microparts precisely, robot calibration, camera calibration and robot-camera registration are needed. Conventional calibration methods are very onerous at the microscale, therefore, two alternative unconventional procedures, called virtual grid calibration and hybrid calibration, are developed for work-cells with high-performance robots, minimising necessary instrumentation. Moreover, an effective calibration of the robot end-effector is designed to compensate for misalignment and orientation errors with respect to the vertical rotational axis. This paper describes the calibration methods and their implementation, the results and the improvements achieved. A detailed comparison between the hybrid and the virtual grid calibrations is provided, demonstrating the higher performance of the latter strategy.

Clonal cultures of Histomonas meleagridis, Tetratrichomonas gallinarum and a Blastocystis sp. were established for the first time. Single microbes were successfully isolated from a mixture of micro-organisms obtained from caecal contents of turkeys, using a micromanipulation approach. The cloned parasites were propagated in vitro and maintained through continuous passages multiplying to high numbers. Identification of the protists was done by morphological investigation identifying various forms of each parasite. PCR and partial sequencing of the small subunit rRNA were used to confirm clonality and to determine the relationship of the cloned parasites with known protozoan parasites. The clonal cultures established by this technique will be useful to gain more insight into the biological repertoire of the organisms. In addition, refined infection experiments in different poultry species can now be performed to elucidate the pathological pathways of the respective protozoa.

We present a visual feedback method for closed loop control of automated microassembly. A CAD model based multi-camera visual tracking system that is well suited for flexible automation and assembly of complex 3D geometries was developed. The system is capable of providing six degree-of-freedom pose feedback on the observed micro-components in real-time (30 Hz). Using CAD models of the observed objects, a complete description of the observed scene, including the effect of occlusions, is available and dependence on distinctive visual features such as fudicial marks is avoided.

A small-scale flexure-based gripper was designed for manipulation tasks requiring precision position and force control. The gripper is actuated by a piezoelectric ceramic stack actuator and utilizes strain gages to measure both the gripping force and displacement. The position and force bandwidths were designed for ten Hertz and one hundred Hertz, respectively, in order to afford human-based teleoperative transparency. The gripper serves effectively as a one degree-of-freedom investigation of compliant mechanism design for position and force controlled micromanipulation. Data is presented that characterizes the microgripper performance under both pure position and pure force control, followed by a discussion of the attributes and limitations of flexure-based design.

Optical tweezers, or the single-beam optical gradient force trap, is becoming a major tool in biology for noninvasive micromanipulation on an optical microscope. The principles and practical aspects that influence construction are presented in an introductory primer. Quantitative theories are also reviewed but have yet to supplant user calibration. Various biological applications are summarized, including recent quantitative force and displacement measurements. Finally, tantalizing developments for new, nonimaging microscopy techniques based on optical tweezers are included.