In Vitro Fertilization (IVF) laboratories often carry a penchant to resist change while in the pursuit of maintaining consistency in laboratory conditions. However, implementation of new technology is often critical to expand scientific discoveries and to improve upon prior successes to advance the field. Microfluidic platforms represent a technology that has the potential to revolutionize the fundamental processes of IVF. While the focus of microfluidic application in IVF has centered on embryo culture, the innovative platforms carry tremendous potential to improve other procedural steps and represents a possible paradigm shift in how we handle gametes and embryos. The following review will highlight application of various microfluidic platforms in IVF for use in maturation, manipulation, culture, cryopreservation and non-invasive quality assessment; pointing out new insights gained into functions of sperm, oocytes and embryos. Platform design and function will also be discussed, focusing on limitations, advancements and future refinements that can further aid in their clinical implementation.

Thinking big by thinking small: application of microfluidic technology to improve ART

Cell transfection is a technique wherein foreign genetic molecules are delivered into cells. To elucidate distinct responses during cell genetic modification, methods to achieve transfection at the single-cell level are of great value. Herein, we developed an automated micropipette-based quantitative microinjection technology that can deliver precise amounts of materials into cells. The developed microinjection system achieved precise single-cell microinjection by pre-patterning cells in an array and controlling the amount of substance delivered based on injection pressure and time. The precision of the proposed injection technique was examined by comparing the fluorescence intensities of fluorescent dye droplets with a standard concentration and water droplets with a known injection amount of the dye in oil. Injection of synthetic modified mRNA (modRNA) encoding green fluorescence proteins or a cocktail of plasmids encoding green and red fluorescence proteins into human foreskin fibroblast cells demonstrated that the resulting green fluorescence intensity or green/red fluorescence intensity ratio were well correlated with the amount of genetic material injected into the cells. Single-cell transfection via the developed microinjection technique will be of particular use in cases where cell transfection is challenging and genetically modified of selected cells are desired.

/pdf/single-cell-transfection-through-precise-microinjection-with-5b3rqztrkj.pdf

Single Cell Transfection through Precise Microinjection with Quantitatively Controlled Injection Volumes

Blastocyst microinjections are routinely involved in the process of creating genetically modified mice for biomedical research, but their efficiency is highly dependent on the skills of the operators. As a consequence, much time and resources are required for training microinjection personnel. This situation has been aggravated by the rapid growth of genetic research, which has increased the demand for mutant animals. Therefore, increased productivity and efficiency in this area are highly desired. Here, we pursue these goals through the automation of a previously developed teleoperated blastocyst microinjection system. This included the design of a new system setup to facilitate automation, the definition of rules for automatic microinjections, the implementation of video processing algorithms to extract feedback information from microscope images, and the creation of control algorithms for process automation. Experimentation conducted with this new system and operator assistance during the cells delivery phase demonstrated a 75% microinjection success rate. In addition, implantation of the successfully injected blastocysts resulted in a 53% birth rate and a 20% yield of chimeras. These results proved that the developed system was capable of automatic blastocyst penetration and retraction, demonstrating the success of major steps toward full process automation.

Blastocyst Microinjection Automation

Three-dimensional positioning and orientation of a biological cell under a microscope is an essential step for cell injection and biopsy. The conventional manual cell manipulation and orientation relies on trial and error, with inherent disadvantages such as low efficiency, poor success rate, and inconsistent output. In this paper, we present a system capable of recognizing and adjusting the orientation of a Zebrafish (Danio rerio) embryo in real time with the use of multiple motion control devices. In addition, a custom-made embryo feeding channel and manipulating device was employed to perform cell manipulations in batches. Experimental results demonstrate the system's capability for fast embryo orientation (13 s/cell) with a 97.5% success rate of rotating a cell to a desired orientation about both y - and z -axis of the cell and a low in-plane rotation angle error of 0.5 ${}^{\circ}$ .

Three-Dimensional Cell Rotation With Fluidic Flow-Controlled Cell Manipulating Device

Microengineering/nanoengineering is an emerging field that enables engineering and scientific discoveries in the microworld. As an effective and powerful tool for automation and manipulation at small scales, precision motion measurement by computer microvision is now broadly accepted and widely used in microengineering/nanoengineering. Unlike other measurement methods, the vision-based techniques can intuitively visualize the measuring process with high interactivity, expansibility, and flexibility. This article aims to comprehensively present a survey of microvision-based motion measurement from the collective experience. Working principles of microvision systems are first introduced and described, where the hardware configuration, model calibration, and motion measurement algorithms are systematically summarized. The characteristics and performances of different microvision-based methods are then analyzed and discussed in terms of measurement resolution, range, degree of freedom, efficiency, and error sources. Recent advances of applications empowered by the developed computer microvision-based methods are also presented. The review can be helpful to researchers who engage in the development of microvision-based techniques and provide the recent state and tendency for the research community of vision-based measurement, manipulation, and automation at microscale/nanoscale.

/pdf/a-review-of-computer-microvision-based-precision-motion-y7cm1fyjql.pdf

A Review of Computer Microvision-Based Precision Motion Measurement: Principles, Characteristics, and Applications

Blastocyst microinjections are routinely involved in the process of creating genetically modified mice for biomedical research, but their efficiency is typically poor and highly dependent on the skills of the operators. Here, the goal of increasing the consistency and efficiency rates of these operations was pursued through robotics and automation. Initially, a teleoperated microinjection system was designed and implemented. Then, this system was evaluated under the control of a microinjection expert. The evaluation consisted of a series of blastocyst microinjections, which are presented and analyzed in this paper. The knowledge gained from these experiments was then used to: 1) create a new system setup more appropriate for automating consecutive blastocyst microinjections; 2) derive rules to guide automatic microinjections; and 3) define future research directions and system upgrades to further increase the system performance. Finally, experiments with the encoded algorithm for automatic blastocyst microinjections attested the validity of the derived rules by demonstrating a 75% success rate. The good quality of the microinjections was demonstrated by the 53% birth rate and the 20% yield of chimeras obtained from the implanted blastocysts.

From teleoperated to automatic blastocyst microinjections: Designing a new system from expert-controlled operations

Cell and embryo microinjections, like many other biomanipulation tasks, involve delicate and precise manual control of micropipettes under a microscope. These operations are repeatedly carried out by highly trained personnel, who spend hours peering through the binoculars of the microscope. As a result, the taxing working conditions and the skills of the operators have a significant impact on the results of the operations. To facilitate the training of new operators and to improve the consistency and efficiency rates of microinjections, we have developed a teleoperated system that demonstrated remarkable improvements in performance and skill acquisition levels during blastocyst microinjection tasks. It combined the advantages of easy, controlled and accurate teleoperation with a more ergonomic work environment, resulting in outstanding reduction in operator training period and amateur performances comparable to those of highly trained operators. This new system, which is fully controlled from a remote computer using a game joystick, was evaluated by comparing teleoperated blastocyst microinjections conducted by one expert and two amateurs. Further, these results were compared to those from expert microinjections conducted on a conventional system. The experiments demonstrated fast learning on the new system and high microinjection success rates (around 90%) for all operators after only three microinjection sessions. Microinjection quality on the new teleoperated system also proved to be high. This was assessed by having the embryos develop to term and computing the birth rates. Higher birth rates (up to twice as high) were observed for embryos injected on the novel system than for those injected on the traditional system. These results prove that the incorporation of robotics into biomanipulation improves the efficiency of the operations, and of the operator, and it eliminates the need for extensive training of microinjection personnel.

Experiments with a teleoperated system for improved bio-micromanipulations

478. A HIV-1 Based Cross-Packaging System for FIV Vectors

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Blastocyst Microinjection Automation

From teleoperated to automatic blastocyst microinjections: Designing a new system from expert-controlled operations

Experiments with a teleoperated system for improved bio-micromanipulations

478. A HIV-1 Based Cross-Packaging System for FIV Vectors