• Title/Summary/Keyword: Pipette

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Pipetting Stability and Improvement Test of the Robotic Liquid Handling System Depending on Types of Liquid (용액에 따른 자동분주기의 분주능력 평가와 분주력 향상 실험)

  • Back, Hyangmi;Kim, Youngsan;Yun, Sunhee;Heo, Uisung;Kim, Hosin;Ryu, Hyeonggi;Lee, Guiwon
    • The Korean Journal of Nuclear Medicine Technology
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    • v.20 no.2
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    • pp.62-68
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    • 2016
  • Purpose In a cyclosporine experiment using a robotic liquid handing system has found a deviation of its standard curve and low reproducibility of patients's results. The difference of the test is that methanol is mixed with samples and the extractions are used for the test. Therefore, we assumed that the abnormal test results came from using methanol and conducted this test. In a manual of a robotic liquid handling system mentions that we can choose several setting parameters depending on the viscosity of the liquids being used, the size of the sampling tips and the motor speeds that you elect to use but there's no exact order. This study was undertaken to confirm pipetting ability depending on types of liquids and investigate proper setting parameters for the optimum dispensing ability. Materials and Methods 4types of liquids(water, serum, methanol, PEG 6000(25%)) and $TSH^{125}I$ tracer(515 kBq) are used to confirm pipetting ability. 29 specimens for Cyclosporine test are used to compare results. Prepare 8 plastic tubes for each of the liquids and with multi pipette $400{\mu}l$ of each liquid is dispensed to 8 tubes and $100{\mu}l$ of $TSH^{125}I$ tracer are dispensed to all of the tubes. From the prepared samples, $100{\mu}l$ of liquids are dispensed using a robotic liquid handing system, counted and calculated its CV(%) depending on types of liquids. And then by adjusting several setting parameters(air gap, dispense time, delay time) the change of the CV(%)are calcutated and finds optimum setting parameters. 29 specimens are tested with 3 methods. The first(A) is manual method and the second(B) is used robotic liquid handling system with existing parameters. The third(C) is used robotic liquid handling system with adjusted parameters. Pipetting ability depending on types of liquids is assessed with CV(%). On the basis of (A), patients's test results are compared (A)and(B), (A)and(C) and they are assessed with %RE(%Relative error) and %Diff(%Difference). Results The CV(%) of the CPM depending on liquid types were water 0.88, serum 0.95, methanol 10.22 and PEG 0.68. As expected dispensing of methanol using a liquid handling system was the problem and others were good. The methanol's dispensing were conducted by adjusting several setting parameters. When transport air gap 0 was adjusted to 2 and 5, CV(%) were 20.16, 12.54 and when system air gap 0 was adjusted to 2 and 5, CV(%) were 8.94, 1.36. When adjusted to system air gap 2, transport air gap 2 was 12.96 and adjusted to system air gap 5, Transport air gap 5 was 1.33. When dispense speed was adjusted 300 to 100, CV(%) was 13.32 and when dispense delay was adjusted 200 to 100 was 13.55. When compared (B) to (A), the result increased 99.44% and %RE was 93.59%. When compared (C-system air gap was adjusted 0 to 5) to (A), the result increased 6.75% and %RE was 5.10%. Conclusion Adjusting speed and delay time of aspiration and dispense was meaningless but changing system air gap was effective. By adjusting several parameters proper value was found and it affected the practical result of the experiment. To optimize the system active efforts are needed through the test and in case of dispensing new types of liquids proper test is required to check the liquid is suitable for using the equipment.

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Analysis of Precipitate Formation Reaction for Measuring Chemical Reaction Rate and Its Development Appling Small-Scale Chemistry (앙금 생성 반응을 이용한 화학반응속도 측정 실험의 분석과 Small-Scale Chemistry를 적용한 실험 개발)

  • Park, Kuk-Tae;Noh, Ji-Hyun;Kim, Dong-Jin;Ryu, Ran-Yeong;Noh, Yun-Mi;Kim, Myo-Kyung;Lee, Sang Kwon
    • Journal of the Korean Chemical Society
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    • v.52 no.3
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    • pp.303-314
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    • 2008
  • The purpose of this study was to understand the experiment for measuring chemical reaction rate by precipitate formation and to develop experiments applying small-scale chemistry. For this study, the experimental method for measuring the effect of concentration and temperature on chemical reaction rates presented in the 10 high school science textbooks were classified by their experimental methods of confirming production. Subsequently, problems observed in carrying out the experiments for measuring chemical reaction rates by precipitate formation frequently presented in the 10 high school science textbooks were analyzed. Experiments applying small-scale chemistry were developed measuring chemical reaction rate by precipitate formation. According to the result of this study, there were some problems in the experimental method of precipitate formation for measuring chemical reaction rates presented in the high school science textbooks. Those problems in the science textbook experiments were insufficient specification of mixing methods of reaction solutions, obscurity of knowing when the character letter X disappeared, time delay in collecting the experimental data, formation of hazardous sulfur dioxide, uneasiness of fixing water bath container, controlling the reaction temperature, and low reproducibility. Those problems were solved by developing experiments applying smallscale chemistry. Presenting the procedure of mixing reaction solutions on the A4 reaction paper sheet made the experimental procedure clearly, using well plates and stem pipette shortened the reaction time and made it possible to continuously collect the experimental data. Furthermore, the quantity of hazardous sulfur dioxide was reduced 1/7 times and the time when the character letter X disappeared could be observed clearly. Since experiments for measuring the effect of concentration and temperature on chemical reaction rates could be performed in 30 minutes, the developing experiments applying SSC would help students understand the scientific concepts on the effect of concentration and temperature on chemical reaction rates with enough time for experimental data analysis and discussion.

Single Cell Dissociation Methods for Flow Cytometric Cell Death Analysis of Hypoxia-Ischemia Injured Newborn Rat Pup Brain (저산소성 허혈성 뇌손상이 유발된 신생백서에서 단일세포의 분리)

  • Hwang, Jong Hee;Sung, Dong Kyung;Choi, Chang Won;Kang, Saem;Chang, Yun Sil;Park, Won Soon;Lee, Munhyang
    • Clinical and Experimental Pediatrics
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    • v.48 no.5
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    • pp.545-550
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    • 2005
  • Purpose : Newborn brain tissue has to be dissociated into a single cell suspension for flow cytometric analysis of cell death during hypoxia-ischemia. Thus the development of a method to dissociate cells from the brain tissue with least damage and maintenance of membrane and antigen integrity remains the challenge for the in vivo application of this technique. We evaluated the efficacy of mechanical or enzymatic (collagenase or tryspin) methods of brain tissue disaggregation. Methods : The extent of the damage to the plasma membrane and loss of the characteristics of the membrane induced with each dissociation method was determined by comparing the flow cytometric results labeled with both fluorescent annexin V and propidium iodide of the newborn rat pup brain tissue in the control group (n=10) and in the 48-hour after hypoxia-ischemia group (n=10). Results : In the control group, the cell percentage of damaged, apoptotic and necrotic cells of both hemispheres with the mechanical dissociation method was significantly increased compared to the trypsin or collagenase method. In the 48-hour after hypoxia-ischemia group, the cell percentage of apoptotic and necrotic cells of the right hemisphere with the collagenase method significantly increased, and live cells significantly decreased compared to the left hemisphere, control group. Although the same trend was observed, the extent of alterations made with the trypsin method was significantly less compared to the collagenase method. Conclusion : The dissociation of neonatal brain tissue for flow cytometric analysis with collagenase was most efficacious with the least cell damage and preservation of the plasma membrane characteristics.

Effects of the Artificial Shrinkage and Assisted Hatching Before Vitrification on the Development of the Vitrified Mouse Expanding Blastocysts (유리화동결 전 인공수축과 보조부화술이 융해 후 생쥐 포배아의 발달에 미치는 영향)

  • Jo, Deok-Hyeon;Ko, Gyoung-Rae;Jung, Ji-Hye;Choi, Jong-Ryeol;Joo, Jong-Kil;Lee, Kyu-Sup
    • Clinical and Experimental Reproductive Medicine
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    • v.35 no.4
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    • pp.275-283
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    • 2008
  • Objective: This study was conducted to investigate the effects of the artificial shrinkage and assisted hatching (PZD; patial zona dissetion) before vitrification on the development of vitrified mouse expanding blastocyst. Methods: Mouse 2-cell embryos were collected and cultured in G1.1 and G2.2 to expanding blastocyst. For artificial shrinkage (AS) the micro injection pipette was inserted into blastocoele cavity and blastocoele fluid was aspirated. For assisted hatching (AH) PZD method was used. Control group was -AS/-AH and treatment groups were -AS/+AH, +AS/-AH and +AS/+AH. After AS and AH mouse blastocysts were equillibrated in G10 and G10E20 for 3 mins, respectively, and vitrified in G25E25 after loading on capped pulled-straw. Vitrified mouse blastocysts were thawed and cultured for 24 hrs. The survival and hatching rate was compared among one control and three treatment groups. Results: The survival rates were 99%, 92% in +AS/+AH and +AS/-AH groups and 54%, 58% in -AS/-AH and -AS/+AH group, respectively. The survival rate was significantly higher in +AS group than in -AS group (p<0.01). Hatching rates were 34%, 96% in -AS/-AH and -AS/+AH groups and 41%, 100% in +AS/-AH and +AS/+AH, respectively. The hatching rates was higher in +AH group than in -AH group (p<0.01). After thawing recovery rates were 100%. Loading on capped pulled-straw, that is effective and useful method on vitrification. Conclusion: This study showed that artificial shrinkage of blastocoele cavity and assisted hatching (PZD) significantly improved the development of the vitrified mouse expanding blastocysts.

Cryopreservation of Primordial Germ Cells(PGCs) from Korean Native Chicken(Ogye) Embryos using Commercial Cryoprotectants (상업용 동결보호제를 이용한 한국재래닭(오계) 원시생식세포의 동결 보존)

  • Kim, Hyun;Kim, Dong Hun;Han, Jae Yong;Do, Yoon Jung;Kim, Jae Hwan;Kim, Young Sin;Seong, Hwan Hoo;Ko, Yeoung Gyu;Kim, Sung Woo
    • Korean Journal of Poultry Science
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    • v.40 no.3
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    • pp.163-169
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    • 2013
  • Cryopreservation of poultry semen has been reported, but preservation of female genetic material has not been possible because of the unique anatomical and physiological characteristics of the avian egg. Thus an alternative strategy for conservation of oviparous species of animals must be developed. Recent technological developments for producing germline chimeras by the transfer of primordial germ cells (PGCs) into recipient embryos has enabled the conservation and retrieval of chicken genetic resources in their complete form. In the present study, fertilized eggs were incubated for about 5.5 days to obtain embryos at stage 28. The whole embryo was collected from the germinal gonad using a fine glass micro pipette under a microscope. The PGCs were then purified using MACS method. Two commercially available cryoprotectants (A and B) were used to preserve the PGCs, and EG were used as a control. The average recovery rate of PGCs after thawing was 35.5% and 60.5% with the A and B treatments, respectively. There was no significant difference between B treatments and control, which showed an average recovery rate of 52.8%. However, the recovery rate obtained using A cryoprotectant (35.5%) was significantly lower than using treatment control and B. The average viability of the PGCs after thawing were 77.9% and 77.4% for cryoprotectants A and B, respectively, and the control were was 81.6%. There was no statistically significant difference between the two treatments and control. It was concluded that all of the available cryoprotectants examined in this study could be used for preservation of PGCs from embryos. Further experiments to produce germline chimera from PGCs preserved using this techniques are strongly recommended.

Biological Characteristics and Growth of the Korean Freshwater Rotifer, Brachionus calyciflorus at Various Temperatures (한국 담수산 윤층 Brachionus calyciflorus의 생물학적 특징과 온도별 성장)

  • 강언종;이배익;김응오
    • Journal of Aquaculture
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    • v.10 no.4
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    • pp.449-456
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    • 1997
  • This study was concuted to determine the optimal conditions for raising the freshwater rotifer, Brachinus calyciflorus. The authors presented some biological informatin obtained from incubation experiment under the various controlled temperatures. Lorica size of the rotifer was divided into two groups : the length and the width for the S-type was $141.0\pm16.7\mu m$($110.1-182.5\;\mu m, n=44$)and $107.0\pm20.3\mu m\;(75.3-152.3\mu m, n=44)$, and those for the L-type was $262.8\pm15.2\mu m\;(234.4-288.6\mu m,\;n=20)\;and\;182.6\pm13.4\mu m (159.8-207.0\mu m,\;n=20$), respectively. The number of eggs being attached on the female varied from 1 to 11 at various culture conditions. Egg type was divided into two groups, large and small. Large and small egg was measured in its major axis as 85a.7-107.8$\mu$m and 55.1-65.2$\mu$m for S-type, and 104.9-121.8 $\mu$m and 62.8-89.1$\mu$m for L-type respectively. The maximum density was reached at 4th day after incubation. The density was 583.9 rotifers/$m\ell$ for $25^{\circ}C$-experimental. group and 421.3 rotifers/$m\ell$ for $22^{\circ}C$-experimental. group respectively. In the case of $28^{\circ}C$-experimental. group, it suddenly decreased into 4.7 rotifers/$m\ell$ at 1st day after incubations and did not recover to its initial density. The maximum rate of increase of populatin per day was reached 0.802 for $22^{\circ}C$-experimental. group at day 2 and fluctuated thereafter. For $25^{\circ}C$-experimental. group it increased to 0.964 at day 3 of incubation and then declined. And the egg ratio of female was reached the maximum of 0.614 for 22$^{\circ}C$- at 3rd day and 0.772 for $25^{\circ}C$-experimental. group at 4th day of incubation.

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Micromorphological and Mineral Characteristics of the Jang-won Series which have Fragipan in the soil Profile (경반층 토양인 장원통의 미세형태학적 및 광물학적 특성)

  • Moon, Yong-Hee;Zhang, Yong-Seon;Chun, Hyen-Chung;Sonn, Yeon-Kyu;Hyun, Byung-Keun;Park, Chan-Won;Song, Kwan-Cheol
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.5
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    • pp.916-921
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    • 2011
  • This study was carry out on a Jang-Won series (fine loamy, mixed, mesic family of typic fragipan) that were established and classified as a fragipan soil in Korea. The morphological, physical, chemical and minerals characteristics of Jang-Won series were studied to determine the genesis of fragipan soils in natural environment. Each sample was analyzed for its physical, chemical and mineralogical characteristics. The particle size distribution of samples was measured using pipette method. Clay minerals were investigated on parallel-oriented specimens of the clay fraction ($<2{\mu}m$) from each horizon, separated by sieving and centrifugation, using X-ray diffraction (XRD) analysis. Micromorphological observations were made on thin sections prepared from soil blocks impregnated with Crystic Resin, cut and ground to less than $30{\mu}m$ in thickness, and finally polished with diamond paste. Most horizons have pH values in the range of fewer than 5.0 and have very low base-saturation values. Their textural classification ranges from silt loam to loam, the lower horizons being the finer. The clay fraction revealed the occurrence of illite, kaolinite, chlorite and vermiculite. The micro-morphological analysis carries out thin sections from each soil profile. The silt concentrations occur as extremely dense and homogenous bands or zones of silt-sized materials, brownish in colour in plane-polarized light and anisotropic in cross-polarized light, surrounding or adhering to skeleton grains. The genesis of fragipan in the Jangweon series assumed composition of clay fraction rather than silt concentration. Therefore, this results suggested an authentic interpretation which Jangweon series is classification as Typic Fragiochrepts.

Distribution Patterns of Surface Sediments of the Jangan Linear Sand Ridge off the Northern Taean Peninsula, in the Mid-west Coast of Korea (서해 중부 태안반도 북부 해역의 장안사퇴 표층퇴적물 분포 특성)

  • TAE SOO CHANG;EUNIL LEE;DO-SEONG BYUN;HWAYOUNG LEE;SEUNG-GYUN BAEK
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.29 no.1
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    • pp.14-27
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    • 2024
  • Unlike the shelf sand ridges moribund in motion, nearshore sand ridges are highly mobile, sensitive to changes in ocean environments, thereby becoming of particular interest with respect to morphological changes. About 5 km off the Daesan port, the Jangan Sand Ridge has been undergoing severe subsea morphological change over the past two decades. Understanding the nature of sand ridges is critical to elucidate the causes of morphological changes. In this context, this study aims at understanding the characteristics and distribution patterns of surface sediments of the ridge and its vicinity. For this purpose, 227 sediment samples were acquired using a grab-sampler, the grain sizes being analysed by the sieve-pipette method. In addition, comparison of grain sizes in sediments between 1997 and 2021 was made in order to investigate the 25-years change in sediment composition. Surface sediments along the ridge axis are fine to medium sands with 2-3 phi in mean grain size, whereas, in the trough of ridge, the sediments are composed of gravels and muddy sandy gravels with mean sizes of -2 to -6 phi. Sediments in the crest of the ridge are well-sorted with normal distribution, on the other hand, the basal sediments are poorly-sorted and positively skewed. Along the ridge crest, the sediments are negatively skewed. From 1997 to 2021, the ridge sediments became largely coarser about 0.5 phi. Such coarsening trend in mean grain size can be explained either by elimination of fine sediments during high waves in winter or elimination of fines suspended during sand mining activities in the past. Spatial distribution pattern of surface sediments shows that ca. 30 m thick of the sand ridge itself overlies the thin relict gravels. The strong asymmetry of sand ridge, the exposure of ridge base, and reworked gravel lags suggest that Jangan sand ridge is probably sediment-deficit and hence erosive in nature at present.

Mineralogy and Biogeochemistry of Intertidal Flat Sediment, Muan, Chonnam, Korea (전남 무안 갯벌 퇴적물에 관한 광물학적 및 생지화학적 연구)

  • Park, Byung-No;Lee, Je-Hyun;Oh, Jong-Min;Lee, Seuug-Hee;Han, Ji-Hee;Kim, Yu-Mi;Seo, Hyun-Hee;Roh, Yul
    • Journal of the Mineralogical Society of Korea
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    • v.20 no.1 s.51
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    • pp.47-60
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    • 2007
  • While sedimentological researches on Western coastal tidal flats of Korea have been much pelformed previously, mineralogical and biogeochemical studies are beginning to be studied. The objectives of this study were to investigate mineralogical characteritics of the inter-tidal flat sediments and to explore phase transformation of iron(oxyhydr)oxides and biomineralization by metal-reducing bacteria enriched from the inter-tidal flat sediments from Muan, Jeollanam-do, Korea. Inter-tidal flat sediment samples were collected in Chungkye-myun and Haeje-myun, Muan-gun, Jeollanam-do. Particle size analyses were performed using the pipette method and sedimentation method. The separates including sand, silt and clay fractions were examined by scanning electron microscopy (SEM) with energy dispersive X-ray (EDX) analysis, transmission electron microscopy (TEM), and X-ray diffiaction (XRD). After enriching the metal-.educing bacteria from the into,-tidal flat sediments, the bacteria were used to study phase transformation of the synthesized iron (oxyhydr)oxides and iron biomineralization using lactate or glucose as the electron donors and Fe(III)-containing iron oxides as the electron accepters. Mineralogical studies showed that the sediments of tidal flats in Chung]rye-myun and Haeje-myun consist of quartz, plagioclase, microcline, biotite, kaolinite and illite. Biogeochemical researches showed that the metal-reducing bacteria enriched from the inter-tidal flat sediments reduced reddish brown akaganeite and mineralized nanometer-sized black magnetite. The bacteria also reduced the reddish brown ferrihydrite into black amorphous phases and reduced the yellowish goethite into greenish with formation of nm-sized phases. These results indicate that microbial Fe(III) reduction may play one of important roles in iron and carbon biogeochemistry as well as iron biomineralization in subsurface environments.