“…The throughput of most existing LoC-based assays is restricted, however, as only a limited number of pollen tubes could be incorporated, guided, and observed on the chip at a time. There have been attempts at LoC-based systems for mechanical characterization of pollen tubes, but they also suffer from low-throughput [14,17] and their closed-cell architecture does not allow interfacing to calibrated micro-indentation [5], micro-gripping [18,19], micro-injection [20], or nano-indentation [21] systems for quantitative biomechanical characterization of the cell wall and cytoplasm.…”
Pollen tubes are used as a model in the study of plant morphogenesis, cellular differentiation, cell wall biochemistry, biomechanics, and intra- and intercellular signaling. For a “systems-understanding” of the bio-chemo-mechanics of tip-polarized growth in pollen tubes, the need for a versatile, experimental assay platform for quantitative data collection and analysis is critical. We introduce a Lab-on-a-Chip (LoC) concept for high-throughput pollen germination and pollen tube guidance for parallelized optical and mechanical measurements. The LoC localizes a large number of growing pollen tubes on a single plane of focus with unidirectional tip-growth, enabling high-resolution quantitative microscopy. This species-independent LoC platform can be integrated with micro-/nano-indentation systems, such as the cellular force microscope (CFM) or the atomic force microscope (AFM), allowing for rapid measurements of cell wall stiffness of growing tubes. As a demonstrative example, we show the growth and directional guidance of hundreds of lily (Lilium longiflorum) and Arabidopsis (Arabidopsis thaliana) pollen tubes on a single LoC microscopy slide. Combining the LoC with the CFM, we characterized the cell wall stiffness of lily pollen tubes. Using the stiffness statistics and finite-element-method (FEM)-based approaches, we computed an effective range of the linear elastic moduli of the cell wall spanning the variability space of physiological parameters including internal turgor, cell wall thickness, and tube diameter. We propose the LoC device as a versatile and high-throughput phenomics platform for plant reproductive and development biology using the pollen tube as a model.
“…The throughput of most existing LoC-based assays is restricted, however, as only a limited number of pollen tubes could be incorporated, guided, and observed on the chip at a time. There have been attempts at LoC-based systems for mechanical characterization of pollen tubes, but they also suffer from low-throughput [14,17] and their closed-cell architecture does not allow interfacing to calibrated micro-indentation [5], micro-gripping [18,19], micro-injection [20], or nano-indentation [21] systems for quantitative biomechanical characterization of the cell wall and cytoplasm.…”
Pollen tubes are used as a model in the study of plant morphogenesis, cellular differentiation, cell wall biochemistry, biomechanics, and intra- and intercellular signaling. For a “systems-understanding” of the bio-chemo-mechanics of tip-polarized growth in pollen tubes, the need for a versatile, experimental assay platform for quantitative data collection and analysis is critical. We introduce a Lab-on-a-Chip (LoC) concept for high-throughput pollen germination and pollen tube guidance for parallelized optical and mechanical measurements. The LoC localizes a large number of growing pollen tubes on a single plane of focus with unidirectional tip-growth, enabling high-resolution quantitative microscopy. This species-independent LoC platform can be integrated with micro-/nano-indentation systems, such as the cellular force microscope (CFM) or the atomic force microscope (AFM), allowing for rapid measurements of cell wall stiffness of growing tubes. As a demonstrative example, we show the growth and directional guidance of hundreds of lily (Lilium longiflorum) and Arabidopsis (Arabidopsis thaliana) pollen tubes on a single LoC microscopy slide. Combining the LoC with the CFM, we characterized the cell wall stiffness of lily pollen tubes. Using the stiffness statistics and finite-element-method (FEM)-based approaches, we computed an effective range of the linear elastic moduli of the cell wall spanning the variability space of physiological parameters including internal turgor, cell wall thickness, and tube diameter. We propose the LoC device as a versatile and high-throughput phenomics platform for plant reproductive and development biology using the pollen tube as a model.
“…Compliant mechanisms (CMs) have been used in a variety of applications at micro and macro scales, such as bioengineering (Bhargav et al, 2015), micro electro mechanical systems (MEMSs) (Zhang et al, 2016;Chen and Ma, 2015;, aerospace (S. L. , laser communication (Clark et al, 2016;Cui et al, 2021), and other fields (Yu et al, 2014;Qiu et al, 2018;Qi et al, 2018). They transmit motion/loads by the elastic deformation of materials.…”
Abstract. In the aerospace field, the precision and stiffness for 2R1T (R denotes the rotation and the T denotes the translation) degree of freedom (DOF) space posture adjustment mechanisms are required. Compliant parallel mechanisms (CPMs) with a suitable constrained branch (SCB) have the advantages of high precision and high stiffness. Based on screw theory, a new type synthesis approach for a 2R1T compliant parallel mechanism with a suitable constrained branch is proposed. The proposed approach is an improvement of the freedom and constraint topology approach. It combines with other methods, including the rigid-body-replacement method, the principle of symmetry, etc. In order to obtain CPMs with a suitable constrained branch, the criterion for the type synthesis is presented. Using this proposed type synthesis approach, a series of CPMs is obtained. They include, but are not limited to, the existing typical 2R1T CPMs with a suitable constrained branch. Furthermore, it identifies the correctness and effectiveness of the approach by analyzing the DOF of the synthesized mechanism. This approach is also suitable for the type synthesis of 4, 5, and 6 DOF compliant parallel mechanisms with a suitable constrained branch.
“…The field of compliant mechanisms is currently being explored extensively in the area of biology, where beambased mechanisms with varying scales are made and are used to manipulate micron level samples. [9,10] A compliant mechanism typically utilizes a flexible single-piece structure to transfer motion or a force from an actuator through elastic deformation of the flexible structure. Another important feature is the scalability of the compliant mechanism for a given application.…”
Objectives:
To demonstrate a novel, non-pneumatic, compliant mechanism-based micro gripper to immobilize oocytes for the Intracytoplasmic sperm injection (ICSI).
Material and Methods:
The micro gripper is designed intuitively based on different techniques available to design compliant mechanisms in the literature such as the Stiffness Maps technique, Kinetoelastostatic maps, and feasibility maps techniques. The gripper’s holder was made from a 2mm thick PMMA sheet; whereas, the gripper was fabricated using a hydrophilic sheet, a proprietary material of 3MTM. The gripper and holder were assembled using a biocompatible adhesive.
Results:
Experimental trials carried out with the gripper holder on the ICSI workbench showed that the developed gripper holder was able to hold the oocyte gently and firmly in place. A micro linear actuator was used to actuate the gripper-holder. The device was tested for its efficacy to perform ICSI by designing ICSI experiments with matured oocytes and sperm; it was found that the degeneration rate was absolutely zero percent for all the matured oocytes.
Conclusions:
A novel device to micro manipulate oocytes during intra-cytoplasmic sperm injection is presented in this paper and is based on the gripping principle as opposed to the conventional suction-based pipettes for holding the oocytes gently and firmly in place during ICSI. The degeneration rate was found to be zero using the gripper-based novel device.
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