Robotic Precisely Oocyte Blind Enucleation Method

Zhao, Xiangfei; Cui, Maosheng; Zhang, Yidi; Liu, Yaowei; Zhao, Xin

doi:10.3390/app11041850

Cited by 6 publications

(3 citation statements)

References 34 publications

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“…However, the process suffers from low efficiency and reproducibility. Manual methods require >1 min per cell (not including locating or retrieving the sample) and suffer from high variability (standard deviation of ±30% of original total volume removed) and low viability (∼40%) . While efforts have been made to use robotically controlled micropipettes and/or computer vision to automate and enhance processes such as enucleation, − microfluidic methods offer a unique advantage in facilitating high precision and high throughput enucleation procedures.…”

Section: Sectioningmentioning

confidence: 99%

Microfluidic Surgery in Single Cells and Multicellular Systems

et al. 2022

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Microscale surgery on single cells and small organisms has enabled major advances in fundamental biology and in engineering biological systems. Examples of applications range from wound healing and regeneration studies to the generation of hybridoma to produce monoclonal antibodies. Even today, these surgical operations are often performed manually, but they are labor intensive and lack reproducibility. Microfluidics has emerged as a powerful technology to control and manipulate cells and multicellular systems at the micro- and nanoscale with high precision. Here, we review the physical and chemical mechanisms of microscale surgery and the corresponding design principles, applications, and implementations in microfluidic systems. We consider four types of surgical operations: (1) sectioning, which splits a biological entity into multiple parts, (2) ablation, which destroys part of an entity, (3) biopsy, which extracts materials from within a living cell, and (4) fusion, which joins multiple entities into one. For each type of surgery, we summarize the motivating applications and the microfluidic devices developed. Throughout this review, we highlight existing challenges and opportunities. We hope that this review will inspire scientists and engineers to continue to explore and improve microfluidic surgical methods.

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Section: Sectioningmentioning

confidence: 99%

Microfluidic Surgery in Single Cells and Multicellular Systems

et al. 2022

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“…The oocyte penetration experiments were performed on the NK-MR601 micromanipulation system [22][23][24], as shown in Figure 1. The system consists of an inverted microscope (CK-40, Olympus, Tokyo, Japan), a CCD camera (W-V-460, Panasonic, Osaka, Japan, frame rate: 20 frames/s), a motorized X-Y stage (travel range: 100 mm, repeatability: 1 µm/s, maximum speed: 2 mm/s), and two X-Y-Z micro-manipulators (travel range: 50 mm, repeatability: 1 µm/s, maximum speed: 1 mm/s); a micro-injector provides the negative pressure to hold the oocyte (−3~0 kPa); a motion control box controls the motorized stage, motorized micro-manipulators, and micro-injector through the host computer.…”

Section: System Setupmentioning

confidence: 99%

Oocyte Penetration Speed Optimization Based on Intracellular Strain

et al. 2022

Self Cite

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Oocyte penetration is an essential step for many biological technologies, such as animal cloning, embryo microinjection, and intracytoplasmic sperm injection (ICSI). Although the success rate of robotic cell penetration is very high now, the development potential of oocytes after penetration has not been significantly improved compared with manual operation. In this paper, we optimized the oocyte penetration speed based on the intracellular strain. We firstly analyzed the intracellular strain at different penetration speeds and performed the penetration experiments on porcine oocytes. Secondly, we studied the cell development potential after penetration at different penetration speeds. The statistical results showed that the percentage of large intracellular strain decreased by 80% and the maximum and average intracellular strain decreased by 25–38% at the penetration speed of 50 μm/s compared to at 10 μm/s. Experiment results showed that the cleavage rates of the oocytes after penetration increased from 65.56% to 86.36%, as the penetration speed increased from 10 to 50 μm/s. Finally, we verified the gene expression of oocytes after penetration at different speeds. The experimental results showed that the totipotency and antiapoptotic genes of oocytes were significantly higher after penetration at the speed of 50 μm/s, which verified the effectiveness of the optimization method at the gene level.

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“…Manual methods require >1 min per cell (not including locating or retrieving the sample) and suffer from high variability (standard deviation of ±30% of original total volume removed) and low viability (~40%). 153 While efforts have been made to use robotically controlled micropipettes and/or computer vision to automate and enhance processes such as enucleation, [153][154][155] microfluidic methods offer a unique advantage in facilitating high precision and high throughput enucleation procedures. The first approach of enucleation is to use solid tools to separate and pinch off the nucleus from the oocyte mechanically inside a microfluidic channel.…”

Section: Enucleationmentioning

confidence: 99%

Microfluidic Surgery in Single Cells and Multicellular Systems

Zhang¹,

Nadkarni²,

Paul³

et al. 2021

Preprint

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Microscale surgery on single cells and small organisms have enabled major advances in fundamental biology and in engineering biological systems. Examples of applications range from wound healing and regeneration studies to the generation of hybridoma to produce monoclonal antibodies. Even today, these surgical operations are often performed manually, but they are labor-intensive and lack reproducibility. Microfluidics has emerged as a powerful technology to control and manipulate cells and multicellular systems at the micro- and nanoscale with high precision. Here, we review the physical and chemical mechanisms of microscale surgery, and the corresponding design principles, applications, and implementations in microfluidic systems. We consider four types of surgical operations: 1) Sectioning, which splits a biological entity into multiple parts, 2) ablation, which destroys part of an entity, 3) biopsy, which extracts materials from within a living cell, and 4) fusion, which joins multiple entities into one. For each type of surgery, we summarize the motivating applications and the microfluidic devices developed. Throughout the review, we highlight existing challenges and opportunities. We hope that this review will inspire scientists and engineers to continue to explore and improve microfluidic surgical methods.

show abstract

Robotic Precisely Oocyte Blind Enucleation Method

Cited by 6 publications

References 34 publications

Microfluidic Surgery in Single Cells and Multicellular Systems

Microfluidic Surgery in Single Cells and Multicellular Systems

Oocyte Penetration Speed Optimization Based on Intracellular Strain

Microfluidic Surgery in Single Cells and Multicellular Systems

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