• Title/Summary/Keyword: Confocal laser scanning microscope

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Expression of Flagellin Proteins of Campylobacter jejuni within Microaerobic and Aerobic Exposures

  • LEE , YOUNG-DUCK;CHOI, JUNG-PIL;MOK, CHUL-KYOON;JI, GEUN-EOK;KIM, HAE-YEONG;NOH, BONG-SOO;PARK, JONG-HYUN
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1227-1231
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    • 2004
  • Campylobacter, one of the emerging foodborne pathogens, is highly adaptable to the external environments by changing its morphology. In the present study, a question of whether the whole-cell antibody would still be effective for its detection even though the morphology of C. jejuni was changed was examined. When microaerophilic C. jejuni was exposed to aerobic conditions for 48 h, its morphological change was detected by confocal laser scanning microscope: Its morphology was confirmed as a spiral-bacilli form in microaerobic condition, however, as a coccoid form with a little spiral-bacilli form, when exposed to aerobic conditions. Also, the expressions of the whole-cell proteins of C. jejuni, and the suppression or induction of newly synthesized proteins in both aerobic and microaerobic conditions were analyzed by two dimensional gel electrophoresis. Additionally, immunoblotting assay with the whole cell antibody for the proteins expressed under the two conditions was performed. It was confirmed that the commercial whole-cell antibody of C. jejuni raised in rabbit was reactive. When analyzed with MALDI- TOF MS, the expressed proteins were confirmed as flagellins. Therefore, even though the morphology changed in aerobic condition, these flagellins were expressed and worked as the eitope proteins, thus making it possible to utilize for the development of an immunosensor for real-time detection of any kind of C. jejuni cell.

Sperm Injection into Maturing and Activated Porcine Oocytes

  • Kim, Bong-Ki;Lee, Yun-Jung;Cui, Xiang-Shun;Kim, Nam-Hyung
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.41-41
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    • 2001
  • Chromatin configuration and microtubule assembly were determined in porcine maturing and activated oocytes following intracytoplasmic sperm injection. Microtubule localization was confirmed using a mouse monoclonal antibody to $\alpha$-tubulin and detected using a fluorescent labeled goat anti-mouse secondary antibody. DNA was stained with propidium iodide. The image of microtubules and chromatin was captured using laser scanning confocal microscope. In germinal vesicle stage oocyte, sperm chromatin remained condensation and sperm derived microtubules were not observed at 8 to 12 h after sperm injection. At 24 h after injection, the sperm nucleus developed to the metaphase chromatin along the metaphase structure of female nucleus. In some metaphase I stage oocytes, sperm chromatin decondensed at 8 h to 12 h after injection, sperm aster was seen soon after sperm injection. At 24 h after sperm injection into metaphase I stage oocyte, male chromatin developed to the metaphase chromatin while female chromatin extruded first polar body and formed the metaphase chromatin. At 12 to 15 h after sperm injection into preactivated oocytes, condensed sperm nucleus was located in close proximity of female pronucleus. However, the condensed nucleus did not fuse with female pronucleus. In preactivated ocytes, injected sperm remained condensation, a few sperm organized small microtubular aster. Instead, maternal derived microtubules were organized near the female chromatin, which seem to move condensed male chromatin near to the female pronucleus. These results suggest that sperm nuclear decondensing activity and nucleation activity of centrosome during fertilization are cell cycle dependent. In absence of male functional centrosome, female origin centrosome takes over the role of microtubule nucleation for nuclear movement.

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Microtubule Assemblies and Methylation Patterns of Porcine IVF and Parthenogenetic Embryos (돼지 체외수정란 및 단위발생란의 미세관 형성 및 메틸화 양상)

  • Park, Joo-Hee;Kim, Ho-Jeong;Lee, Beom-Ki;Kwon, Dae-Jin;Hwang, In-Sun;Park, Choon-Keun;Yang, Boo-Keun;Cheong, Hee-Tae
    • Reproductive and Developmental Biology
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    • v.33 no.1
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    • pp.7-11
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    • 2009
  • This study was performed to confirm the microtubule assemblies and methylation patterns of porcine IVF and parthenogenetic embryos. Cumulus-oocyte complexes were collected and matured in vitro for 42 hr. Oocytes were fertilized by prepared fresh sperm or activated parthenogenetically by exposure to electric stimulation and 6-dimethylaminopurine. Porcine IVF and parthenogenetic embryos were cultured in vitro for 6 days. Embryos were stained by immunofluorescence staining method to observe the dynamic of nucleus and microtubules in the first mitotic phase and the methylation patterns in different developmental stages. After then, samples were confirmed and analyzed through a laser-scanning confocal microscope. IVF embryos had a centrosome originated from sperms, which was shown a $\gamma$-tubulin spot. However, $\gamma$-tubulin spot was not observed in parthenogenetic embryos. A lower methylation level was observed in IVF embryos compared to parthenogenetic ones at the morula and blastocyst stages. In conclusion, it is considered that microtubule assemblies and genetic regulation mechanism differ between parthenogenetic and IVF embryos.

Distribution of Glutamate Receptors in the Retina of the Greater Horseshoe Bat (Rhinolophus ferrumequinum) (한국관박쥐 망막에서 글루타메이트 수용체의 분포 양상)

  • Kwon, Oh-Ju;Jeon, Chang-Jin
    • Journal of Korean Ophthalmic Optics Society
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    • v.19 no.3
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    • pp.413-418
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    • 2014
  • Purpose: The objective of this study was analyzing the distribution of the excitatory neurotransmitter glutamate receptor to investigate the function in the retina of the greater horseshoe bat. Methods: After retinal tissues of adult greater horseshoe bat were cut into $40{\mu}m$ vertical sections, standard immuno-cytochemical techniques was applied for analysis. Immunofluorescence images were obtained using the Bio-Rad MRC 1024 laser scanning confocal microscope. Results: AMPA (GluR1-4), Kainate (GluR5-7, KA1-2) and NMDA (1, 2A, 2B) mainly distributed in the inner plexiform layer and outer plexiform layer. KA1 receptors have existed not only plexiform layer but also ganglion cell layer. Conclusions: The greater horseshoe bat has same neuron and neurotransmitter to mammalian retina. These findings suggest that bat has a functional retina for visual analysis.

Cariogenicity of Vitamin Supplements for Children (시판용 어린이 비타민 보충 제제의 치아 우식원성)

  • No, Yoomi;Kim, Jongsoo;Yoo, Seunghoon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.45 no.2
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    • pp.195-202
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    • 2018
  • The purpose of this study is to analyze the cariogenicity of vitamin supplements for children by the Caries Potentiality Index (CPI), pH drop capacity, proliferation rate of Streptococcus mutans. Four vitamin supplements were selected - Noma (NM), Cenovis Kids (CK), Animal Parade (AP), and Character Vitamin (CV). CPI value decreased in the order of AP, CV, CK, and NM. Initial values of all experimental groups showed acidity below pH 7.0. Analysis of the colony forming units of Streptococcus mutans showed that NM and CV resulted a higher proliferation rate (p < 0.05) than CK and AP (p < 0.05). Bacterial activity of the control group was lower than other groups when observed with a confocal laser scanning microscope. Considering the bacterial activity and acidity of vitamin supplements, it is necessary to pay close attention when children taken the vitamin supplements for their oral health.

Effects of Fiber Wall Thickness on Paper Properties Using CLSM (CLSM을 이용한 고해과정 중 섬유벽 두께 변화의 종이 특성 영향 분석)

  • 김서환;박종문;김철환
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.31 no.1
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    • pp.39-45
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    • 1999
  • Refining in papermaking plays an important role in changing fiber properties as well as paper properties. The major effects of refining on pulp fibers are internal and external fibrillation, fiber shortening, and fines formation. Many workers showed that internal fibrillation of the primary refining effects was most influential in improving paper properties. In particular, refining produces separation of fiber walls into several lamellae, thus causing fiber wall swelling with water penetration. This leads to the increase of fiber flexibility and of fiber-to-fiber contact during drying. If the fibers are very flexible, they will be drawn into close contact with each other by the force of surface tension as the water is removed during the drainage process and drying stages. In order to study the effect of fiber wall delamination on paper properties, cross-sectional image of fibers in a natural condition had to be generated without distortion. Finally, it was well recognized that confocal laser scanning microscope (CLSM) could be one of the most efficient tool for creating and quantifying fiber wall delamination in combination with image analysis technique. In this study, the CLSM could be used not only to observe morphological features of transverse views of swollen fibers refined under low and high intensity, but also to investigate the sequence of fiber wall delamination and fiber wall breakage. From the CLSM images, increasing the specific energy or refining decreased the degree of fiber collapse, fiber cross-sectional area, fiber wall thickness and lumen area. High intensity refining produced more external fibrillation.

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Effect of NaCl on Biofilm Formation of the Isolate from Staphylococcus aureus Outbreak Linked to Ham

  • Lee, Soomin;Choi, Kyoung-Hee;Yoon, Yohan
    • Food Science of Animal Resources
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    • v.34 no.2
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    • pp.257-261
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    • 2014
  • The objective of this study was to evaluate the effects of NaCl on the biofilm formations of the isolate from Staphylococcus aureus outbreaks linked to ham. The S. aureus ATCC13565 isolated from ham was exposed to NaCl concentrations of 0%, 2%, 4%, and 6% supplemented in tryptic soy broth (TSB) for 24 h at $35^{\circ}C$, followed by plating 0.1 mL of the culture on tryptic soy agar containing 0%, 2%, 4%, and 6% NaCl, respectively. After incubating at $35^{\circ}C$ for 24 h, the colonies on the plates were collected and diluted to $OD_{600}$ = 0.1. The diluents of S. aureus were incubated on a 96-well flat bottom plate containing TSB plus the appropriate NaCl concentrations, and the biofilm formation was quantified by crystal violet staining after being incubated at $35^{\circ}C$ for 9 h. Confocal laser scanning microscope (CLSM) was also used for visualizing the biofilm formation of S. aureus at NaCl concentrations of 0%, 2%, 4%, and 6%. The transcriptional analysis of biofilm-related genes, such as icaA, atl, clfA, fnbA, sarA, and rbf, was conducted by quantitative real-time PCR. Crystal violet staining and CLSM showed that the biofilm formations of S. aureus increased (p<0.05) along with the NaCl concentrations. Moreover, the expression of the icaA genes was higher at the NaCl concentrations of 4% and 6% as compared with 0% of NaCl by approximately 9-folds and 20-folds, respectively. These results indicated that the NaCl formulated in processed food may increase the biofilm formations of S. aureus by increasing the icaA gene expressions.

The Influence of Microinjection of Foreign Gene into the Pronucleus of Fertilized Egg on the Preimplantation Development, Cell Number and Diameter of Rabbit Embryos

  • Makarevich, A.V.;Chrenek, P.;Fl’ak, P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.2
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    • pp.171-175
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    • 2006
  • The aim of this in vitro study was to test the effect of microinjection (Mi) of foreign gene into the rabbit egg pronucleus and epidermal growth factor (EGF) addition on the blastocyst rate, the cell number and the diameter of embryos, and to determine possible relationships between embryo cell number and embryo diameter. Blastocyst rate was significantly decreased in gene- Mi (G-Mi/E0) group (63.1%) comparing to intact ones (83.5%, $p_1$<0.05). The addition of EGF at 20ng/ml (G-Mi/E20) or 200 ng/ml (GMi/ E200) to gene-Mi embryos did not affect blastocyst rate (65.6 and 55.2% resp.). As a control for Mi, the eggs were microinjected with the same volume of phosphate-buffered solution (PBS-Mi) instead of the gene construct solution. Cell numbers and embryo diameters were measured from embryo images obtained on confocal laser scanning microscope. Bonferroni-modified LSD test showed that the embryo cell number in PBS-Mi group was significantly lower ($p_1$<0.05) and in gene-Mi group was tended to decrease compared with intact embryos. Embryo diameter was not different among experimental groups. No effect of EGF given at any doses both on the cell number and embryo diameter was found. A positive correlation between cell number and embryo diameter was observed in all groups of embryos. Since embryo diameter was not changed under the influence of Mi or EGF addition in this study, this seems to be more conservative characteristics of the embryo morphology. These results suggest that the pronuclear microinjection compromises developmental potential of embryos, decreasing blastocyst rate and embryo cell number, whilst embryo diameter is not affected. No effects of EGF on studied parameters were confirmed. Declined quality of Mi-derived embryos is caused by the microinjection procedure itself, rather than by the gene construct used.

CLSM Analysis of Change in Roughness and Physical Properties of Granite after Freeze-Thaw Experiments (CLSM을 이용한 동결/융해 실험 후 화강암 시료의 표면 및 물성변화 분석)

  • Jeong, Jongtaek;Choi, Junghae;Chae, Byung-Gon;Jang, Bo-An
    • The Journal of Engineering Geology
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    • v.23 no.3
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    • pp.271-281
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    • 2013
  • Freeze-thaw experiments were conducted to evaluate changes in surface roughness and physical properties in samples of granite from Ilgwang and Imki mines, Korea. The temperature range in the experiments was $-20^{\circ}C$ to $40^{\circ}C$, based on typical summer and winter temperatures in Korea, and the surface was observed every 20 cycles. One cycle comprised 1 hour of heating or cooling of the samples and 1 hour during which the target temperature was maintained. With increasing repetitions of the freeze-thaw experiment, porosity increased by 0.05%-0.15% in the two samples and the dry weight increased, whereas the volume of the soil and saturation weight decreased. Observations by confocal laser scanning microscope (CLSM) revealed that line and surface roughness parameters showed a tendency to increase and decrease, respectively, with elapsed time. Changes in surface roughness were apparent on the CLSM images.

In Situ Observation of Solidification Behavior in Undercooled $Pd_{40}Cu_{30}Ni_{10}P_{20}$ Alloy Melts during Linear Cooling (연속냉각 중 과냉 된 $Pd_{40}Cu_{30}Ni_{10}P_{20}$ 합금 용탕의 실시간 응고거동 관찰)

  • Kim, Ji-Hun
    • Journal of Korea Foundry Society
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    • v.23 no.5
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    • pp.276-285
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    • 2003
  • In the undercooled melt of $Pd_{40}Cu_{30}Ni_{10}P_{20}$ alloy, the solidification behavior including nucleation and growth of crystals at the micrometer level has been observed in-situ by use of a confocal scanning laser microscope combined with an infrared image furnace. The $Pd_{40}Cu_{30}Ni_{10}P_{20}$ alloy specimens were cooled from the liquid state to glass transition temperature. 575 K, at various cooling late under a helium gas flow. According to the cooling rate, the morphologies of the solidification front are changed among various types, irregular jog like front, columnar dendritic front, cellular grain, star like shape jog and fine grain, etc. The velocities of the solid-liquid interface are measured to be $10^{-5}{\sim}10^{-8}$ m/s which are at least two orders higher than the theoretical crystal growth rates. Combining the morphologies observed in terms of cooling rates and their solidification behaviors, we conclude that phase separation takes place in the undercooled molten $Pd_{40}Cu_{30}Ni_{10}P_{20}$ alloy. The continuous cooling transformation (CCT) diagram was constructed from solidification onset time at various linear cooling conditions with different rate. The CCT diagram suggests that the critical cooling rate for glassy solidification is about 1.5 K/s, which is in agreement with the previous calorimetric findings.