• Journal of Embryo Transfer
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• v.14 no.2
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• pp.107-111
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• 1999

The present study was performed to evaluate the best electric fusion condition in nuclear transfer, Korean Native Cattle fibroblasts were used as nucleic donors. Oocytes from slaughterhouse were matured in vitro for 22 h and enucleated. Each individual cells were transferred into enucleated ocytes and reconstructed embryo were placed into the fusion chamber. In experiment 1, pulse were performed by altering pulse duration at 1. 75kv/cm, 1 time. When pulse duration is 30$mutextrm{s}$, fusion and development rates is higher than other conditions. In experiment 2, the effect of different pulse number were studied at the pulse duration 30$mutextrm{s}$ and the same pulse intensity. When pulse number was one, fusion rates were higher than other conditions. The fused embryos were moved to culture medium and assessed their development to blastocyst. These results showed that best fusion condition was 30$mutextrm{s}$ and one time. And the fibroblasts derived from Han Woo can be reprogrammed by nuclear transplantation and develop subsequently in vitro.

• Korean Journal of Animal Reproduction
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• v.18 no.3
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• pp.217-228
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• 1994

These experiments were carried out to establish the optimal condition of electrostimulatin inducing cell fusion and oocyte activation for nuclear transplantation in mouse embryos. Eggs selected for cell fusion or activation by electrostimulation were equilibrated for 5~10 min. in 0.3M sucrose solution and electrostimulated for 60$\mu$sec using 1 pulse of 60, 70, 80, 90 or 100 volts DC with electrodes 0.2 mm apart. Then they were cultured in 20${mu}ell$ dropsof Tyrode's solution. The results of these experiments are as follows : 1. When one pulse of 60, 70, 80, 90 or 100 volts DC for 60$\mu$sec were applied to 2-cell embryos for fusion of blastomeres, fusion rates were 50.0, 81.7, 91.7, 100 and 100%, respectively ; and developmental rates of fused embryos to blastocyst were 76.7 to 81.5%. Higher fusion rates were observed in 90V and 100V. 2. The average cell number in fused embryos developed to blastocyst was about half of the cell number in diploid controls; and the cell number decreased with increasing of voltages. 3. When pulse numbers were increased, fusion rates improved, but developmental rates were not signficiantly different from the group for which the number of pulse was not increased. And the cell number of blastocyst decreased even more. 4. Oocytes aged for 6hrs after ovulation were electrostimulated for oocyte activation by the same method used for cell fusion. Rates of oocyte activated by electrostimulation were 45.3 to 60.4%, and fragmentation rates were 7.5~15.1%. The lysis rates were 17.0~34.0%. The results of these experiments indicate that the optimal condition for achieving cell fusion and activation is 1 pulse, duration 60$\mu$sec in 90 Volt. The results also show that this condition is suitable for nuclear transplantation using mouse eggs.

• Proceedings of the Korean Nuclear Society Conference
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• 2004.10a
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• pp.719-720
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• 2004

A multi-megawatt long pulse ion source (LPIS) of neutral beam injector was developed for the KSTAR. Beam extraction experiments of the LPIS were carried out at the neutral beam test stand (NBTS). Design requirements for the ion source were 120 kV/65 A deuterium beam and a 300 s pulse length. A maximum ion density of $9.1310^{11}$ $cm^{-3}$ was measured by using electric probes, and an optimum arc efficiency of 0.46 A/kW was estimated with ion saturation current of the probes, arc power, and total beam area. An arcing problem, caused by the structural defect of decelerating grid supporter, in the third gap was solved by the blocking of backstream ion particles, originated from the plasma in the neutralizer duct, through the unnecessary spaces on the side of grid supporter. A maximum drain power of 1.5 MW (i.e. 70 kV/21 A) with hydrogen was measured for a pulse duration of 0.5 s. Optimum beam perveance was ranged from 0.75 to 0.85. An improved design of accelerator for the effective control of beam particle trajectory should provide higher beam perveance.

• Journal of Biomedical Engineering Research
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• v.15 no.1
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• pp.1-8
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• 1994

Electrofusion of cabbage protoplasts was performed with a developed equipment which consists of a parallel wire electrode, a AC field generator, and a pulse generator. Exposure of protoplasts to an alternating current field of 450 KHz causes attraction to each other which leads to chains of protoplasts extending from the electrode. Intra-specific protoplast fusion was effectively induced within the pearl chains by the additional application of a single high-intensity DC square wave pulse of 1 KV/cm for 1-3 msec. To improve the performance, micro fusion electrode fabricated from the semiconductor technology and microcontroller based field an, d pulse generator was proposed. The viability of protoplasts after an application of electric field at optimum condition was reduced by only 5 % compared with that of pre-application. A prototype of fusion electrode, which consists of insulator-structure, was successfully fabricated by using photosensitive polyimide.

• Korean Journal of Animal Reproduction
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• v.19 no.3
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• pp.171-179
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• 1995

This study was carried out to investigate the fertilizing ability of round spematids isolated from seminiferous tubules. A round spermatid was introduced into the perivitelline space of a mature oocyte using Leitz micromanipulators and then subjected to electrofusion. Electrofusion was induced by applying a single DC pulse of 90V with a duration of 60$\mu$sec using Model 611 Square Wave Stimulator(Phipps and Bird, U.S.A) in 0.3 M sucrose fusion medium containing 0.05mM CaCl2 and 0.1mM MgSO4, Oocyte pre-activation was conducted by exposure to a single DC(80V, 80$\mu$sec) pulse in electrofusion medium at 1 hour before electrofusion. The incidence of fusion with pre-activated oocytes(23.8%, 57/239) was higher than that with nonactivated oocytes(6.7%, 3/45). The most of electro-stimulated mouse oocytes cleaved regardless of the success or failure of fusion. Karyotyping of embryos that developed into blastocysts after exposure to the fusion pulse were performe. We found that blastocysts from the fused oocytes were diploid whereas blastocysts from the unfused oocytes were haploid. About 11.7 and 11.5% of fused and unfused oocytes were developmental potentials of fused and unfused oocytes. Therefore, these results suggest that the mouse mture oocyte can be fertilized by fusion with a round spermtid and subsequently developed normally.

• Proceedings of the Korean Nuclear Society Conference
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• 2005.10a
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• pp.887-888
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• 2005

• Journal of Biomedical Engineering Research
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• v.17 no.2
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• pp.173-178
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• 1996

The objective of this paper is to investigate the effect of AC field on the protoplast of plant cells. The results of investigation will be the basis for the development of etectric cell fusion device. For the experiment, we made the electrode and AC and DC pulse generator and observed the behavior of the protoplasts through the inverted microscope which is connected to the monitor and video recorder by the CCD camera. As a result, the numbers of rotating, moving and destructed protoplasts and viability of the protoplasts have close relation to the amplitude of AC field, while the rotation rate is closely related to the frequency of AC pulse.

• Korean Journal of Animal Reproduction
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• v.24 no.4
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• pp.365-373
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• 2000

The purpose of this study was to investigate the effects of the fusion pulses and fusion media on fusion rate and the development of embryos produced by somatic cell nuclear transfer in Hanwoo (Korean cattle). Nuclear donor cumulus and fetal fibroblast cells were cultured in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at 38.5$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$in air. The in vitro matured oocytes were enucleated and then the isolated donor cells were introduced. The cumulus cell and cytoplast were fused using one pulse of 70 volts for 40$mutextrm{s}$, two pulses of 70 volts for 40$mutextrm{s}$ and one pulse of 180 volts for 15$mutextrm{s}$. The fetal fibroblast cell and cytoplast were fused using one pulse of 180 volts for 15$mutextrm{s}$ or 30$mutextrm{s}$. The cumulus cell and cytoplast were fused using mannitol and Zimmerman cell fusion medium (ZCFM) as a fusion medium. The fused embryos were activated after the fusion with 10 $\mu$M calcium ionophore for 5 min and 2 mM 6-dimethyl- aminopurine for 3 h. The nuclear transfer embryos were cultured in 500 ${mu}ell$ well of modified CR1aa supplemented with 3 mg/$m\ell$ BSA in th $\varepsilon$ four well dish cove red with mineral oil. After 3 days culture, culture medium was changed into modified CRlaa medium containing 1.5 mg/$m\ell$ BSA and 5% FBS for 4 days. The incubation environment was 5% $CO_2$, 5% $O_2$, 90% $N_2$ at 38.5$^{\circ}C$. When the cumulus cells were fused with enucleated oocytes by three different fusion pulses, one pulse of 180 volts for 15 $mutextrm{s}$ yielded the highest fusion rate and developmental rate to blastocyst among the pulses (P＜0.05). When the fetal fibroblast cells were fused with enucleated oocytes, one pulse of 180 volts for 30$mutextrm{s}$ yielded significantly higher fusion rate compared with that for 15 $mutextrm{s}$(P＜0.05). The present result indicates that the fusion rate between karyoplast and cytoplast was affected by the cell type and the optimal fusion condition was different according to cell type or size. When the fusion was conducted by the use of mannitol and ZCFM, the fusion rate was 71.2% and 65.8%, respectively. The developmental rates to blastocyst were 37.8% and 39.8%, respectively. There was no significant difference between two fusion media in the developmental rate of cumulus cell nuclear transfer embryos. These results indicate that optimal electric current should be selected according to cell type.

• KSII Transactions on Internet and Information Systems (TIIS)
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• v.14 no.11
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• pp.4395-4412
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• 2020

The multisource image fusion has become an active topic in the last few years owing to its higher segmentation rate. To enhance the accuracy of multimodal pig-body feature segmentation, a multisource image fusion method was employed. Nevertheless, the conventional multisource image fusion methods can not extract superior contrast and abundant details of fused image. To superior segment shape feature and detect temperature feature, a new multisource image fusion method was presented and entitled as NSST-GF-IPCNN. Firstly, the multisource images were resolved into a range of multiscale and multidirectional subbands by Nonsubsampled Shearlet Transform (NSST). Then, to superior describe fine-scale texture and edge information, even-symmetrical Gabor filter and Improved Pulse Coupled Neural Network (IPCNN) were used to fuse low and high-frequency subbands, respectively. Next, the fused coefficients were reconstructed into a fusion image using inverse NSST. Finally, the shape feature was extracted using automatic threshold algorithm and optimized using morphological operation. Nevertheless, the highest temperature of pig-body was gained in view of segmentation results. Experiments revealed that the presented fusion algorithm was able to realize 2.102-4.066% higher average accuracy rate than the traditional algorithms and also enhanced efficiency.

• Proceedings of the Korean Vacuum Society Conference
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• 2011.02a
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• pp.240-240
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• 2011

The first neutral beam injector (NBI-1) has been developed for the Korea Superconducting Tokamak Advanced Research (KSTAR) tokamak. A first long pulse ion source (LPIS-1) has been installed on the NBI-1 for an auxiliary heating and current drive of KSTAR core plasmas. Performance of ion and neutral beam extractions in the LPIS-1 was investigated initially on the KSTAR NBI-1 system, prior to the neutral beam injection into the main plasmas. The ion source consists of a JAEA magnetic bucket plasma generator with multi-pole cusp fields and a set of KAERI prototype-III tetrode accelerators with circular apertures. The inner volume of plasma generator and accelerator column in the LPIS-1 is approximately 123 liters. Final design requirements for the ion source were a 120 kV/ 65 A deuterium beam and a 300 s pulse length. The extraction of ion beams was initiated by the formation of arc plasmas in the LPIS-1, called as an arc-beam extraction method. A stable ion beam extraction of LPIS-1 has been achieved up to an 100 kV/42 A for a 4 s pulse length and an 80 kV/25 A for a 14 s pulse length. Optimum beam perveance of 1.21 microperv has been found at an accelerating voltage of 80 kV. Neutralization efficiency has been measured by using a water flow calorimetry (WFC) method of calorimeter and an operation of bending magnet. The full-energy species of ion beams have been detected by using the diagnostic method of optical multichannel analyzer (OMA). An arc efficiency of the LPIS was 0.6~1.1 A/kW depending on the operating conditions of arc discharge.