• Applied Microscopy
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• v.43 no.1
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• pp.14-20
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• 2013

The leopard danio, Danio rerio var. frankei is a spotted color morph of the zebrafish, Danio rerio caused by a pigment mutation. The structural differences of fertilized egg and egg envelope are poorly documented. To clarify this, we compared the fertilized egg morphology and ultrastructures of surface structures, the micropyle and the cross section of fertilized egg envelopes of zebrafish and leopard danio, variation species of zebrafish using a light and electron microscopes. Although the fertilized egg sizes were different, the external shapes of the fertilized eggs of two species couldn't be differentiated under the light microscope. The characteristics of fertilized eggs, such as a spherical shape, a non-adhesive quality and a large perivitelline space, were shown to be related to spawning habit. In ultrastructure of fertilized egg envelope, there is no morphological difference of micropyle between two species. By contrast, the ultrastructure and the numbers of knob-like structures and semihemisphere-like structures per unit area on the outer surface, and the number of lamellae of inner layer on the fertilized egg envelope section displayed definite species specificity. Collectively, our data indicate that the ultrastructure of fertilized egg envelope in the zebrafish could be differentiated by species variation.

• 제어로봇시스템학회:학술대회논문집
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• 2003.10a
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• pp.2451-2456
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• 2003

In biological cell manipulation, manual thrust or penetration of an injection pipette into an egg is currently performed by a skilled operator, relying only on visual feedback information. Massive load of various micro injection of either genes, fluid or cells in the postgenomic era calls a more reliable and automatic micro injection system that can test hundreds of genes or cell types at a single experiment. We initiated to study cellular force sensing in zebrafish eggs as the first step for the development of a more controllable micro injection system by any inexperienced operator. Zebrafish eggs at different developmental stages were collected and an integrated biomanipulation system was employed to measure cellular force during penetrating the egg envelope, the chorion. First of all, the biomanipulation system integrated with cellular force sensing instrument is implemented to measure the penetration force of cell membranes and characterize mechanical properties of zebrafish embryo cells. Furthermore, implementation of cellular force sensing system and calibration are presented. Finally, the cellular force sensing of penetrating cell membranes at each developmental stages was experimentally performed. The results demonstrated that the biomanipulation system with force sensing capability can measure cellular force at real-time while the injection operation is undergoing. The magnitude of the measured force was in the range of several hundreds of uN. The precise real-time measurement should provide the first step forwards for the development of an automatic and reliable injection system of various materials into biological cells.

• 제어로봇시스템학회:학술대회논문집
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• 2003.10a
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• pp.2457-2460
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• 2003

There is a great demand to manipulate biological cell autonomously since biologist should spend much time to obtain skillful manipulation techniques. For this purpose, we propose a cell chip to control, carry, fix and locate the cell. In this paper, we focus on the cell rotator to rotate individual biological cell based on a micro fluidics technology. The cell rotator consists of injection hole and rotation well to rotate a biological cell properly. Under the variation of flow rate in injection hole, the angular velocity of a biological cell is evaluated to find the feasibility of the proposed rotation method. As a practical experiment, Zebrafish egg is employed. Based on this research, we find the possibility of non-contact rotation way that can highly reduce the damage of the biological cell during manipulation. To realize an autonomous biological cell manipulation, a cell chip with manipulation well and micro channel in this research will be utilized effectively in near future.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.283-283
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• 2004

The eggs of most animal species are surrounded by an extracellular matrix known as chorion, egg envelope, egg coat, or zona pellucida. Development of fish embryo usually takes several days in an aquatic environment. During embryonic development, the chorion must protect embryo from physical damage and microbial infection in the exposed aquatic environment. (omitted)

• Transactions of the Korean Society of Mechanical Engineers A
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• v.27 no.12
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• pp.2079-2084
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• 2003

In biological cell manipulation, manual thrust or penetration of an injection pipette into an embryo cell is currently performed by a skilled operator, relying on visual feedback information only. Accurately measuring cellular forces is a requirement for minimally invasive cell injections. Moreover, the cellular force sensing is essential in investigating the biophysical properties for cell injury and membrane modeling studies. This paper presents cellular force measurements for the force feedback-based biomanipulation. Cellular force measurement system using piezoelectric polymer sensor is implemented to measure the penetration force of a zebrafish egg cell. First, measurement system setup and calibration are described. Second, the force feedback-based biomanipulation is experimentally carried out. Experimental results show that it successfully supplies real-time cellular force feedback to the operator at tens of uN and thus plays a main role in improving the reliability of biological cell injection tasks.

• Proceedings of the Korean Society of Precision Engineering Conference
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• 2003.06a
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• pp.237-240
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• 2003

In biological cell manipulation, manual thrust or penetration of an injection pipette into an embryo cell is currently performed by a skilled operator, relying on visual feedback information only. Accurately measuring cellular forces is a requirement for minimally invasive cell injections. Moreover, the cellular farce sensing is essential in investigating the biophysical properties for cell injury and membrane modeling studies. This paper presents cellular force measurements for the force feedback-based biomanipulation. Cellular force measurement system using piezoelectric polymer sensor is implemented to measure the penetration force of a zebrafish egg cell. First, measurement system setup and calibration are described. Second, the force feedback-based biomanipulation is experimentally carried out. Experimental results show that it successfully supplies real-time cellular force feedback to the operator at several tens of uN and thus plays a main role in improving the reliability of biological cell injection tasks.