• Title/Summary/Keyword: Cryo

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The Effect of Various Vitrification Methods on Developmental Rate of Mouse Pronuclear Embryos at Different Recovery Times (다양한 유리화 동결 방법이 각 시간대별 생쥐 전핵기 배아의 발달에 미치는 영향)

  • Kim, Ji-Chul;Seo, Byoung-Boo;Park, Sung-Baek;Kim, Jae-Myeoung
    • Journal of Embryo Transfer
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    • v.27 no.1
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    • pp.63-69
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    • 2012
  • The objective of this study was to investigate the effectiveness of cryopreservation methods for the effect of various vitrification containers, such as EM-grid, OPS, or cryo-loop on the survival and developmental rate of vitrified mouse pronuclear embryos, and mouse cleavage embryo, at 21, 24, 27 and 30 hr after hCG injection. Post-thaw cleavage was similar among treatments, while the developmental rates of mouse blastocyst and hatched blastocyst were higher ($p$ <0.05) in 27 hr and 30 hr than 21 hr. The developmental rate of hatched blastocyst at vitrified cleavage mouse embryos in cryo-loop was significantly higher than vitrified pronuclear embryos of control group as well as EM-grid and OPS ($p$ <0.05). The developmental rate using cryo-loop was higher than EM-grid, but in case of OPS at vitrified cleavage and mouse pronuclear embryos, no significant difference was noticed. These results of our study show that the developmental rates of mouse embryos were unaffected by various vitrification containers, but in case of mouse embryos and hatched blastocysts at late vitrified pronuclear embryos the developmental rates were higher than early vitrified pronuclear embryos. Moreover, the developmental rate of hatched blastocyst at vitrified cleavage mouse embryos was significantly higher than vitrified pronuclear embryos. For better execution of this study, it will be mandatory to include improvement of vitrification containers, cryopreservation methods and conditions, higher survival rate, safe preservation, contamination and embryo loss.

Purification of wastewater from paper factory by cryo-cooled high-$T_c$ superconducting magnetic separator (전도냉각형 고온초전도 자석을 이용한 제지폐수의 자기분리에 의한 정수)

  • Ha, Dong-Woo;Kim, Tae-Hyung;Sohn, Myung-Hwan;Kwon, Jun-Mo;Baik, Seung-Kyu;Oh, Sang-Soo;Ko, Rock-Kil;Ha, Hong-Soo;Kim, Ho-Sup;Kim, Young-Hun;Kang, Che-Hun;Ha, Tae-Wook
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2009.05a
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    • pp.4-4
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    • 2009
  • Paper factories use a large amount of water and same amount of wastewater is generated. It is important to recycle the wastewater because of water shortages and water pollution. The existing water treatment facilities like precipitation process need large-scale equipment and wide space to purify the wastewater of paper factory. High gradient magnetic separation (HGMS) system has the merits to purify rapidly because of large voids at filter and to occupy small space. In this paper, two types of superconducting magnets were used for HGMS systems. Cryo-cooled Bi-2223 superconducting magnet system with 70 mm room temperature bore and 200 mm of height was prepared. Cryo-cooled Nb-Ti superconducting magnet with 100 mm room temperature bore and 600 mm of height was used for magnetic separator. Magnetic filters were designed by the analysis of magnetic field distribution at superconducting magnets. The various magnetic seeding reactions were investigated to increase the reactivity of coagulation. The effects of magnetic separation of wastewater were investigated as variation of magnetic field strength and flow rate of wastewater.

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Effect of Temperature and Surfactant on Crystallization of Al-Based Metallic Glass during Pulverization (분쇄 공정의 온도와 분산제 사용이 알루미늄계 금속유리의 결정화에 미치는 영향)

  • Tae Yang Kim;Chae Yoon Im;Suk Jun Kim
    • Korean Journal of Materials Research
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    • v.33 no.2
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    • pp.63-70
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    • 2023
  • In this study, crystallization was effectively suppressed in Al-based metallic glasses (Al-MGs) during pulverization by cryo-milling by applying an extremely low processing temperature and using a surfactant. Before Al-MGs can be used as an additive in Ag paste for solar cells, the particle sizes of the Al-MGs must be reduced by milling. However, during the ball milling process crystallization of the Al-MG is a problem. Once the Al-MG is crystallized, they no longer exhibit glass-like behavior, such as thermoplastic deformation, which is critical to decrease the electrical resistance of the Ag electrode. The main reason for crystallization during the ball milling process is the heat generated by collisions between the particles and the balls, or between the particles. Once the heat reaches the crystallization temperature of the Al-MGs, they start crystallization. Another reason for the crystallization is agglomeration of the particles. If the initially fed particles become severely agglomerated, they coalesce instead of being pulverized during the milling. The coalesced particles experience more collisions and finally crystallize. In this study, the heat generated during milling was suppressed by using cryo-milling with liquid-nitrogen, which was regularly fed into the milling jar. Also, the MG powders were dispersed using a surfactant before milling, so that the problem of agglomeration was resolved. Cryo-milling with the surfactant led to D50 = 10 um after 6 h milling, and we finally achieved a specific contact resistance of 0.22 mΩcm2 and electrical resistivity of 2.81 μΩcm using the milled MG particles.

Copper Oxide Spike Grids for Enhanced Solution Transfer in Cryogenic Electron Microscopy

  • Dukwon Lee;Hansol, Lee;Jinwook Lee;Soung-Hun Roh;Nam-Chul Ha
    • Molecules and Cells
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    • v.46 no.9
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    • pp.538-544
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    • 2023
  • The formation of uniform vitreous ice is a crucial step in the preparation of samples for cryogenic electron microscopy (cryo-EM). Despite the rapid technological progress in EM, controlling the thickness of vitreous ice on sample grids with reproducibility remains a major obstacle to obtaining high-quality data in cryo-EM imaging. The commonly employed classical blotting process faces the problem of excess water that cannot be absorbed by the filter paper, resulting in the formation of thick and heterogeneous ice. In this study, we propose a novel approach that combines the recently developed nanowire self-wicking technique with the classical blotting method to effectively control the thickness and homogeneity of vitrified ice. With simple procedures, we generated a copper oxide spike (COS) grid by inducing COSs on commercially available copper grids, which can effectively remove excess water during the blotting procedure without damaging the holey carbon membrane. The ice thickness could be controlled with good reproducibility compared to non-oxidized grids. Incorporated into other EM techniques, our new modification method is an effective option for obtaining high-quality data during cryo-EM imaging.

Investigation on Effective Operational Temperature of HTS Cable System considering Critical Current and AC loss

  • Kim, Tae-Min;Yim, Seong-Woo;Sohn, Song-Ho;Lim, Ji-Hyun;Han, Sang-Chul;Ryu, Kyung-Woo;Yang, Hyung-Suk
    • KEPCO Journal on Electric Power and Energy
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    • v.2 no.2
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    • pp.307-310
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    • 2016
  • The operational cost for maintaining the superconductivity of high-temperature superconducting (HTS) cables needs to be reduced for feasible operation. It depends on factors such as AC loss and heat transfer from the outside. Effective operation requires design optimization and suitable operational conditions. Generally, it is known that critical currents increase and AC losses decrease as the operational temperature of liquid nitrogen ($LN_2$) is lowered. However, the cryo-cooler consumes more power to lower the temperature. To determine the effective operational temperature of the HTS cable while considering the critical current and AC loss, critical currents of the HTS cable conductor were measured under various temperature conditions using sub-cooled $LN_2$ by Stirling cryo-cooler. Next, AC losses were measured under the same conditions and their variations were analyzed. We used the results to select suitable operating conditions while considering the cryo-cooler's power consumption. We then recommended the effective operating temperature for the HTS cable system installed in an actual power grid in KEPCO's 154/22.9 kV transformer substation.

Development of an Efficient Processor for SIRAL SARIn Mode

  • Lee, Dong-Taek;Jung, Hyung-Sup;Yoon, Geun-Won
    • Korean Journal of Remote Sensing
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    • v.26 no.3
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    • pp.335-346
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    • 2010
  • Recently, ESA (European Space Agency) has launched CryoSAT-2 for polar ice observations. CryoSAT-2 is equipped with a SIRAL (SAR/interferometric radar altimeter), which is a high spatial resolution radar altimeter. Conventional altimeters cannot measure a precise three-dimensional ground position because of the large footprint diameter, while SIRAL altimeter system accomplishes a precise three-dimensional ground positioning by means of interferometric synthetic aperture radar technique. In this study, we developed an efficient SIRAL SARIn mode processing technique to measure a precise three-dimensional ground position. We first simulated SIRAL SARIn RAW data for the ideal target by assuming the flat Earth and linear flight track, and second accessed the precision of three-dimensional geopositioning achieved by the proposed algorithm. The proposed algorithm consists of 1) azimuth processing that determines the squint angle from Doppler centroid, and 2) range processing that estimates the look angle from interferometric phase. In the ideal case, the precisions of look and squint angles achieved by the proposed algorithm were about -2.0 ${\mu}deg$ and 98.0 ${\mu}deg$, respectively, and the three-dimensional geopositioning accuracy was about 1.23 m, -0.02 m, and -0.30 m in X, Y and Z directions, respectively. This means that the SIRAL SARIn mode processing technique enables to measure the three-dimensional ground position with the precision of several meters.

Development of In Vitro Produced Bovine Embryos after Vitrification with Various Containers (다양한 Container로 유리화 동결된 체외생산 소 수정란의 발달에 관한 연구)

  • Lee, Y.J.;Ko, D.H.;H.T., Lee;Chung, K.S.
    • Korean Journal of Animal Reproduction
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    • v.25 no.4
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    • pp.371-379
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    • 2001
  • The objective of this study was to establish an effective cryopreservation method of in vitro-produced bovine embryos. For the vitrification, in virtro-produced embryos at 8-cell, morula and blastocyst stages were exposed to freezing solution containing 5.5 M EG (EG 5.5) for 20 sec, loaded on each containers such as EM grid, OPS and Cryo-loop, and then immediately plunged into liquid nitrogen at -196$^{\circ}C$. Thawed embryos were serially diluted in 0.5, 0.25 and 0.125 M sucrose in m-HPBS, each for 1 min, and cultured in CRlaa medium supplemented with 10% FBS. Significant differences in the rates of re-expanded and hatched embryos were not observed among these embryo containers. The total cell number of expanded blastocyst cultured in vitro after vitrification was examined by Hoechst staining. There were no differences between non-vitrified (180.0 $\pm$ 5.4) and vitrified groups (178.0 $\pm$ 7.5). In addition, when the cellular injuries after vitrification were compared by double staining. There were no significant difference in the ratio of live and dead cells between non-vitrified group (176 : 4) and vitrified group (172 : 6). Therefore, these results suggest that bovine embryos can be cryopreserved easily, effectively and successfully by vitrification using various containers, such as EM grid, OPS or Cryo-loop in the presence of EG 5.5 freezing solution.

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Role of $K^+$ Channels in $H_2O_2$- and Cryo-induced Apoptosis of Mouse and Bovine Embryos (과산화수소와 동결에 의해 유도된 생쥐와 소 수정란의 사멸에 있어서 칼륨 통로의 역할)

  • Choe, Changyong;Kim, Chang-Woon;Kang, Dawon;Han, Jaehee
    • Journal of Embryo Transfer
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    • v.29 no.3
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    • pp.249-255
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    • 2014
  • Programmed cell death or apoptosis is associated with changes in $K^+$ concentration in many cell types. Recent studies have demonstrated that two-pore domain $K^+$ ($K_{2P}$) channels are involved in mouse embryonic development and apoptotic volume decrease of mammalian cells. In cerebellar granule neurons that normally undergo apoptosis during the early developmental stage, TASK-1 and TASK-3, members of $K_{2P}$ channels, were found to be critical for cell death. This study was performed to identify the role of $K^+$ channels in the $H_2O_2$-induced or cryo-induced cell death of mouse and bovine embryos. Mouse and bovine two-cell stage embryos (2-cells) exposed to $H_2O_2$ for 4 h suffered from apoptosis. The 2-cells showed positive TUNEL staining. Treatment with high concentration of KCl (25mM) inhibited $H_2O_2$-induced apoptosis of 2-cells by 19%. Cryo-induced death in bovine blastocysts showed positive TUNEL staining only in the cells near the plasma membrane. Cryoprotectant supplemented with 25 mM KCl reduced apoptosis slightly compared to cryoprotectant supplemented with 5 mM KCl. However, the combination of antioxidants (${\beta}$-mercaptoethanol) with 25 mM KCl significantly decreased the rate of $H_2O_2$-induced and cryo-induced apoptosis compared to treatments with only antioxidants or 25 mM KCl. These results show that blockage of $K^+$ channel efflux for a short-time reduces $H_2O_2$- and cryo-induced apoptosis in mouse and bovine embryos. Our findings suggest that apoptosis in mouse and bovine embryos might be controlled by modulation of $K^+$ channels which are highly expressed in a given cell type.

THE EFFICACY OF PROGRAMMED CRYO-PRESERVATION UNDER PRESSURE IN RAT PERIODONTAL LIGAMENT CELLS (압력 저속 냉동 방법의 쥐 치아 치주인대세포 보존 효율 평가)

  • Lee, Young-Eun;Kim, Eui-Seong;Kim, Jin;Han, Seung-Hoon;Lee, Seung-Jong
    • Restorative Dentistry and Endodontics
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    • v.34 no.4
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    • pp.356-363
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    • 2009
  • The purpose of this study was to evaluate the viability of periodontal ligament cells in rat teeth using slow cryo-preservation method under pressure by means of MTT assay and WST-1 assay. Eighteen teeth of Sprague-Dawley white female rats of 4 week-old were used for each group. Both sides of the first and second maxillary molars were extracted as atraumatically as possible under Tiletamine anesthesia. The experimental groups were group 1 (Immediate control), group 2 (Cold preservation at $4^{\circ}C$for 1 week), group 3 (Slow freezing), group 4 (Slow freezing under pressure of 3 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in $37^{\circ}C$water bath, then MTT assay and WST-1 assay were processed. One way ANOVA and Tukey method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization. In both MTT and WST-1 assay, group 4 showed significantly higher viability of periodontal ligament cells than group 2 and 3 (p < 0.05), but showed lower viability than immediate control group. By the results of this study, slow cryo-preservation method under pressure suggests the possibility for long term cryo-preservation of the teeth.

Structural Analysis of Exosomes Using Different Types of Electron Microscopy

  • Choi, Hyosun;Mun, Ji Young
    • Applied Microscopy
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    • v.47 no.3
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    • pp.171-175
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    • 2017
  • Negative staining has been traditionally used for exosome imaging; however, the technique is limited to surface topology only and can cause staining artifacts. Therefore, to analyze the internal structure of exosomes, we employed a method of block preparation, thin sectioning, and electron tomography. In addition, an automatic serial sectioning technique with 15-nm thickness through focused ion beam was employed to observe the three-dimensional structure of exosomes of various sizes. Cryo-transmission electron microscopy revealed the near-to-native structure of exosomes.