• Korean Journal of Animal Reproduction
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• v.19 no.4
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• pp.259-264
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• 1996

This study was carried out to investigate on the survival rates and in vitro developmental rates of bisected bovine embryos were by manipulator and micropipette. Bisected embryos were co-cultured in 20% FCS(v/v)＋TCM-199 media containing hormones, oviductal epithelial cells and cumulus cells 0 to 72 hrs after bisection. Survival rate and in vitro fertilization rate were defined as development rate on in vitro culture or FDA-test. The results are summarized as follows ; 1. The in vitro developmental rate of biseciton embryos co-cultured in 20% FCS＋TCM-199 medium containing PMSG, hCG, PMSG＋hCG, hCG＋$\beta$-estradiol 0 to 20 hrs and 20 to 40 hrs were 36.7, 26.7, 33.3, 40.0, and 30.0, and 30.0, 33.3, 30.0, 26.7, and 26.7%, respectively. The survival rate of bisection embryos co-cultured in TCM-199 medium containing hormones was significantly higher than that of non co-culture(25.0%). 2. The in vitro developmental rates of bisection embryos co-cultured in 20% FCS＋TCM-199 medium containing oviductal epithelial cells 4 to 5 hrs and 20 to 24 hrs were 40.0 and 33.3%, respectively. The survival rate of bisection embryos co-cultured in TCM-199 medium containing oviductal epithelial cells was significantly higher than that of non co-culture(25.0%). 3. The in vitro developmental rates of bisection embryos co-cultured in 20% FCS＋TCM-199 medium containing cumulus cells 4 to 5 hrs and 20 to 24 hrs were 43.3 and 36.7%, respectively. The survival rate of bisection embryos co-cultured in TCM-199 medium containing cumulus cells was significantly higher than that of non co-culture (25.0%).

• Journal of Biosystems Engineering
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• v.35 no.4
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• pp.257-261
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• 2010

Surface energy is the principal factor of anti-condensation. High surface energy appears hydrophilic itself and low surface energy represents hydrophobic itself. The contact angle is widely being used for measurement of surface energy of materials, evaluation of coating performances, measurement of wettability, and so on. However, the existing contact angle measuring system is so expensive for purchasing and complicated, so it takes a lot of time and money to use. This study was conducted to develop the algorithm for evaluating hydrophilic performance through measuring the contact angle of water droplet automatically, and fabricate relatively simple measuring system using a low-cost monochrome camera and image processing. A constant amount of water was firstly allocated on a slide by a micropipette, and then the image of water droplet was captured by monochrome digital camera and sent to a computer. The image was binarized and then reduced noises by labeling. Finally, the contact angle of water droplet was computed by using three points (left, right, and top coordinates), simple linear mathematics, and trigonometric function. The experimental results demonstrated the accuracy and reproducibility of the developed system showing less deviations and deviation ratio.

• Clinical and Experimental Reproductive Medicine
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• v.41 no.2
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• pp.68-74
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• 2014

Objective: In search of an ideal method of assisted hatching (AH), we compared the effects of conventional micropipette-AH and laser-AH on the blastocyst formation rate (BFR) and blastocyst cell numbers. Methods: Four-to five-week-old ICR female mice were paired with male mice after superovulation using Pregnant mare's serum gonadotropin (PMSG) and hCG. The two-cell embryos were flushed from the oviducts of female mice. The retrieved two-cell embryos underwent one of five AH procedures: single mechanical assisted hatching (sMAH); cross mechanical assisted hatching (cMAH); single laser assisted hatching (sLAH); quarter laser assisted hatching (qLAH); and quarter laser zona thinning assisted hatching (qLZT-AH). After 72 hours incubation, double immunofluorescence staining was performed. Results: Following a 72 hours incubation, a higher hatching BFR was observed in the control, sMAH, cMAH, and sLAH groups, compared to those in the qLAH and qLZT-AH groups (p<0.05). The hatched BFR was significantly higher in the qLAH and qLZT-AH groups than in the others (p<0.05 for each group). The inner cell mass (ICM) was higher in the control and sMAH group (p<0.05). The trophectoderm cell number was higher in the cMAH and qLAH groups (p<0.05). Conclusion: Our results showed that the hatched BFR was higher in groups exposed the the qLAH and qLZT-AH methods compared to groups exposed to other AH methods. In the qLAH group, although the total cell number was significantly higher than in controls, the ICM ratio was significantly lower in than controls.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.84-84
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• 2002

The advantages of the OPS techniques(Vajta G et al, Mol Reprod Dev 51: 53-58,1998) give 1) high survival rates of various types of eggs, 2) quick and simple process, 3) inexpensive equipment and reduced chilling injury. The efficiency of IVM/IVF technique in the porcine species is relatively lower than that obtained in other species such as ruminants. Two experiments were designed to investigate the effects of in-vitro fertilization of porcine oocytes matures using different OPS protocol for chilling and warming of vitrification. Porcine oocytes from ovaries collected at abattoir were matured for 44 hours in TCM199 Earle's salt supplemental with pyruvate, pff, L-cysteine, hormones and gentamycin. Oocytes were denuded and fertilized with frozen boar semen by common method. Porcine embryos produced routinely by in-vitro culture system of NCSU23 medium. The vitrification and the warming were conducted by OPS method with the glass micropipette instead of straw vessels and modified the protocol of G.Vajta(1999). In Exp 1, Chilling/Warming:Holding Medium(HM)+EG+DMSO/HM +sucrose Medium(SM) at 39$^{\circ}C$ warm stage. In Exp 2, : PBS+CS+EG+Ficoll+ Trehalose/PBS+Trehalose at 25$^{\circ}C$ stage. Filling, freezing, packing, thawing out and further culturing were performed to follow the basic protocol of G Vajta. During IVM-lVC and post-warming, fertilization parameter and developmental potential were compared to and statistically analysed. It was not significantly different from Exp 1 and Exp 2 but 25$^{\circ}C$ of stage was slightly higher on the morula/blastocyst forming rate and better atmosphere for worker than that at 39$^{\circ}C$ stage.

• Transactions of the Korean Society of Mechanical Engineers B
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• v.35 no.11
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• pp.1133-1137
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• 2011

A novel technique for the fabrication of a glass micropipette-based thermal sensor was developed utilizing inexpensive thermocouple materials. Thermal fluctuation with a resolution of ${\pm}0.002$ K was measured using the fabricated thermal probe. The sensors comprise unleaded low-melting point solder alloy (Sn) as a core metal inside a borosilicate glass pipette coated with a thin film of Ni, creating a thermocouple junction at the tip. The sensor was calibrated using a thermally insulated calibration chamber, the temperature of which can be controlled with a precision of ${\pm}0.1$ K and the thermoelectric power (Seebeck coefficient) of the sensor was recorded from 8.46 to $8.86{\mu}V$/K. The sensor we have produced is both cost-effective and reliable for thermal conductivity measurements of micro-electromechanical systems (MEMS) and biological temperature sensing at the micron level.

• Journal of Embryo Transfer
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• v.12 no.1
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• pp.21-27
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• 1997

The studies were carried out to investigate the effects of bisection method and with and without-zona pellucida of embryos on in vitro developmental rate bisected embryos by micromanipulator, micropipette and pipetting. The ovaries were obtained from slaughtered Korean native cows. The follicular oocytes were cultured in TCM-199 mediurn containing 10 IU /ml의 PMSG(Sigma, USA), 10 IU /ml의 hCG, 1$\mu$g /ml의 $\beta$-estradiol(Sigma, USA) and 10% FCS for 24~48 hrs in incubator with 5% $CO_2$ in air at 38.5$^{\circ}C$ and then, matured oocytes were again cultured for 12~ 18 hrs with motile capacitated sperm by preincubation of heparin. Bisected embryos cultured for 1~5 days in 20% FCS + TCM-199 medium. Survival rate was defined as developmental rate on in vitro culture or FDA-test. The results are summarized as follows :1. The survival rates of bisected bovine embryos by micromanipulator and micropipett were 29.2% and 19.1%, respectively. The rates of non-bisection embryos(46.7%) were significantly higher than those of bisection embryos. 2. The in vitro developmental rates of bisected bovine embryos by micromanipulator, micropipett and pipetting method were 32.4%, 19.4% and 25.6%, respectively.3. The in vitro developmental rates of with and without-zona pellucida of bisected bovine embryos by raicromanipulator were 30.8% and 25.0%, respectively. The rates of nonbisection embryos(53.1%) were significantly higher than those of bisection embryos.

• The Korean Journal of Pain
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• v.32 no.3
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• pp.168-177
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• 2019

Background: Brennan's rodent paw incision model has been extensively used for understanding mechanisms underlying postoperative pain in humans. However, alterations of physiological parameters like blood pressure and heart rate, or even feeding and drinking patterns after the incision have not been documented as yet. Moreover, though eicosanoids like prostaglandins and leukotrienes contribute to inflammation, tissue levels of these inflammatory mediators have never been studied. This work further investigates the antinociceptive effect of protein C after intra-wound administration. Methods: Separate groups of Sprague-Dawley rats were used for quantitation of cyclooxygenase (COX) activity and leukotriene B4 level by enzyme-linked immunosorbent assay, as well as estimation of cardiovascular parameters and feeding and drinking behavior after paw incision. In the next part, rats were subjected to incision and $10{\mu}g$ of protein C was locally administered by a micropipette. Both evoked and non-evoked pain parameters were then estimated. Results: COX, particularly COX-2 activity and leukotriene B4 levels increased after incision. Hemodynamic parameters were normal. Feeding and drinking were affected on days 1 and 3, and on day 1, respectively. Protein C attenuated non-evoked pain behavior alone up to day 2. Conclusions: Based upon current observations, Brennan's rodent paw incision model appears to exhibit a prolonged period of nociception similar to that after surgery, with minimal interference of physiological parameters. Protein C, which is likely converted to activated protein C in the wound, attenuated the guarding score, which probably represents pain at rest after surgery in humans.

• Korean Journal of Animal Reproduction
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• v.25 no.3
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• pp.259-267
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• 2001

This study was carried out to investigate on the improvement of fertilizing and developing ability of in vitro matured oocytes from sperm density, motility and PVP(polyvinylpyrrolidone), HA(hyaluronic acid) concentrations, sperm capacitation and intact, free-zona of bovine oocytes obtained by intracytoplasmic sperm injection(ICSI). 1. The fertilization and cleavage rates of bovine oocytes obtained by ICSI treated 0.01, 0.02, 0.03, 0.05% of PVP concentrations were 72.7∼90.9% and 38.5∼54.5%, respectively and these values of 0.02% addition of PVP were higher than other concentrations of PVP 2. The fertilization and cleavage rates of bovine oocytes obtained by ICSI treated 0.01, 0.02% HA and 0.02% PVP + HA concentrations were 72.7%, 40.9% and 81.8%, 54.5% and 83.3%, 37.5%, respectively. and these values of 0.02% addition of PVP + HA were lowe than other concentrations of HA. 3. The fertilization and cleavage rates of bovine oocytes obtained by ICSI treated fresh and frozen sperm were 93.3%, 85.7% and 60.0%, 46.7%, respectively and these values of fresh sperm injection were higher than frozen sperm. 4. The fertilization and cleavage rates of bovine oocytes obtained by ICSI capacitated sperm of heparin, BFF and His methods were 86.7%, 78.9%, 65.0% and 61.9%, 52.6%, 50.0%, respectively. 5. The fertilization and cleavage rates of zona-intact and zona-free bovine oocytes obtained by ICSI were 84.2%, 78.3% and 57.9%, 34.8%, respectively. 6. The fertilization and cleavage rates of bovine oocytes obtained by IVF or ICSI were 63.3∼64.6%, 88.2∼90.0% and 26.7∼29.2%, 52.9∼67.5%, respectively. This ICSI method improved high fertilization rates of bovine oocytes.

• Korean Journal of Veterinary Research
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• v.38 no.4
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• pp.898-908
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• 1998

The aim of this experiments were to assess the time-interval change of motional characteristics in frozen-thawed semen of Korean native cattle (KNC) by using computer aided semen analysis (CASA) technology. Twenty-six KNC frozen semen straws were obtained from Korean KNC improvement department, livestock improvement main division, national livestock cooperatives federation in Korea. Specimens were allowed to thaw at $37^{\circ}C$ for 30 sec in water bath. Semen analysis was performed on semen image analysis system (SIAS, Medical supply, Korea) adjusted to the gate settings and used the semen droplet ($5{\mu}l$) placed on Makler counting chamber (Sefi medical instrument, Israel) prewarmed at $37^{\circ}C$. The same person used the same micropipette to fill the Makler counting chamber. A total of 150 or more of sperms were analysed in each specimen by a single trained person by scanning at least 5 to 10 fields. The measurement parameters in SIAS were as follows ; frame rate = 30 frames per sec, image capture = 1 sec, minimum motile speed = $10{\mu}m/s$, maximum countable sperm number = 400. Statistical analysis was done by Student t-test with use of the Sigma plot program on a IBM personal computer. The dancemean(DNM) and hyperactivated sperm(HYP) of frozen-thawed KNC semen kinematics were significantly decreased(p < 0.05) after 10 min of incubation at $37^{\circ}C$ water bath. But, wobble(WOB) of same sample semen was significantly increased(p < 0.05) after 10 min of incubation and significantly decrease(p < 0.05) after 60 min of same incubation. And, after 30 mim of incubation, significantly differences were found most of motion kinematics, motifity(MOT), curvilinear velocity(VCL), straight line velocity(VSL), average path velocity(VAP), amplitude of lateral head displacement(ALH), beat cross frequency(BCF), mean angular displacement(MAD), dance(DNC), on same sample semen. The DNM of KNC semen sample was variable kinematics after 30 min of incubation. Also, the linearity(LIN) and straightness(STR) was significantly decreased(p < 0.05) from 60 min of incubation. In conclusion, the AI within 30 min after thawing of frozen semen can be an effective method for obtaining high fertility rate in KNC reproductive program.

• Journal of Marine Bioscience and Biotechnology
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• v.2 no.3
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• pp.174-186
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• 2007

Amphora coffeaeformis and Achnanthes longipes are commonly found as dominant benthic microalgae in Jeju coastal water throughout the year. In order to investigate pharmaceutical uses of these diatoms, each single species was isolated with micropipette under phase contrast microscope and subcultured with synthetic seawater media which was enriched with F/2 media, trace metal solution and $Na_2SiO_3$). Growth characteristics of these species were also determined with different combination of salinity, nutrients concentration and temperature. Thereafter, mass culture of each species was done based on the maximum growth condition. Biomass was collected after two weeks of mass culture and freeze dried for antioxidant study. The antioxidant properties of different fractions (n-hexane, chloroform and ethylacetate) obtained by solvent fractionation of 80% methanolic extract of two microalgae were investigated for free radical, reactive oxygen species scavenging (Super oxide, Hydrogen peroxide, Hydroxyl radical and Nitric oxide), metal chelating and lipid peroxidation inhibition activities. All fractions of A. longipes showed higher $DPPH^{\cdot}$ (free radical) scavenging activities (n-hexane: 89.0%, Chloroform: 76.0%, Ethylacetate: 66.0%, Methanol: 90.6% and aqueous residue: 63.0%). N-hexane fraction of A. longipes showed significantly higher activity (49.0%) on nitric-oxide. Ethylacetate fraction of A. longipes and aqueous residue of A. coffeaeformis exhibited 64.0% and 75.6% metal chelating activity which was higher than commercial antioxidants (${\alpha}$-tocopherol: 18.0% and BHT: 16.0%). The n-hexane fraction of A. coffeaeformis had 67.5% activity on $DPPH^{\cdot}$. Chloroform and n-hexane fractions of A. coffeaeformis exhibited 46.2% and 47.6% $H_2O_2$ scavenging effects which were closely similar to commercial antioxidants (${\alpha}$-tocopherol: 49.2% and BHT: 58.6%). Chloroform and ethylacetate fractions of A. longipes and fraction of n-hexane and chloroform of A. coffeaeformis showed better lipid peroxidation activities than ${\alpha}$-tocopherol. These data suggest that both organic and aqueous fractions have good antioxidative compounds with different antioxidant properties.