• Applied Microscopy
• /
• v.35 no.3
• /
• pp.121-128
• /
• 2005

It has been known that ras signaling transduction leads to cell proliferation and migration including various adaptor molecules. Dynamin protein has been implicated in the formation of nascent vesicles in both the endocytic and secretory pathways. Dynamin was classified into three isoforms: dynamin I is only expressed in neuronal tissue, dynamin II is expressed ubiquitously in all tissue but that of dynamin III is confined to testis. We have reported in previous study that Grb2, binding to ras, was associated with dynamin II in NIH3T3 cells. Therefore we have tried to identify the relative expression of dynamin II according to overexpressed ras protein in ras oncogene transfected cells (NIH3T3 (ras)). For the detection of differential expression of dynamin II, we have used immunofluorescent staining and western blot methods in NIH3T3 and NIH3T3 (ras) cells. Next we have described the morphological differences between NIH3T3 and NIH3T3 (ras) cells using SEM and TEM. From these experiments dynamin II was highly expressed in NIH3T3 (ras) cells. NIH3T3 cells was transformed to more spindle shape with many cell process by transfection of ras oncogene. Moreover dynamin II was more concentrated in endocytotic membrane of the NIH3T3 (ras) cells compared to that of NIH3T3 cells. The present results suggested that dynamin II may involve the intermediate messenger in Ras signaling transduction pathway.

• Journal of the korean academy of Pediatric Dentistry
• /
• v.44 no.2
• /
• pp.170-179
• /
• 2017

The purpose of this study was to compare the cariogenicity of commercially available bovine milk and low-fat milk in a biofilm model using the CDC Biofilm Reactor. Streptococcus mutans ATCC 25175 biofilms were formed on saliva coated bovine enamel slabs in a CDC Biofilm Reactor. Biofilms were exposed three times per day to one of the following materials: commercial whole milk (fat content: 3.4%), low-fat milk (fat content: 1%), or 0.9% NaCl. Medium pH was measured at different time points. After 5 days, biofilms were separated from slabs to evaluate the CFUs. The biofilm thickness was observed by confocal laser-scanning microscopy (CLSM). Enamel slab's demineralization was assessed by measuring surface microhardness before and after the experiment. For microhardness and CFUs assessment, no significant difference was found among the three groups. All groups showed similar pattern of medium pH change and biofilm thickness. Our results showed that there was no difference in the cariogenicity between whole milk and low-fat milk. Both milks were relatively non-cariogenic compared to the control group.

• Journal of Korean Ophthalmic Optics Society
• /
• v.20 no.1
• /
• pp.51-60
• /
• 2015

Purpose: To determine the effect of marketed multipurpose contact lens solutions (MPSs) on human corneal epithelial cells (HCEpiCs) toxicity by using image analysis. Methods: HCEpiCs were exposed six MPSs (product A-F) at 0.05~50% for 2h, 12h, 24h, and 48h respectively. HCEpiCs were fixed and stained with Draq5 after exposure with MPSs, and the cell viability and apoptosis were evaluated by using confocal microscope and ImageXpress UltraTM. Results: Viabilities of HCEpiCs exposed to MPS A-F for a 2h were not affected, while reductions (52~75%) in cell viability over a 12h exposure of MPS B, MPS C, MPS D and MPS F, and significant more reductions (29~73%) over a 24h and 48h-exposure. Apoptosis of HCEpiC was not affect over a 12h MPS exposure, however was significantly increased (199~526%) over 24h and 48h MPS exposure. Among the products MPS D, E and F reduced viability of HCEpiCs and apoptosis increased more than MPS A (p<0.05). Conclusions: Lower concentration of MPSs have not an cytotoxic effect on HCEpiCs, however higher concentration of MPSs induce apoptosis and reduce viability of HCEpiCs. Therefore, it need to develop MPS having antimicrobial effectiveness with low cytotoxicity.

• Proceedings of the Korean Vacuum Society Conference
• /
• 2014.02a
• /
• pp.109-110
• /
• 2014

주사전자현미경(Scanning Electron Microscopy: SEM)은 고체상태에서 미세조직과 형상을 관찰하는 데에 가장 다양하게 쓰이는 분석기기로서 최근에 판매되고 있는 고분해능 SEM은 수 나노미터의 분해능을 가지고 있다. 그리고 SEM의 초점심도가 크기 때문에 3차원적인 영상의 관찰이 용이해서 곡면 혹은 울퉁불퉁한 표면의 영상을 육안으로 관찰하는 것처럼 보여준다. 활용도도 매우 다양해서 금속파면, 광물과 화석, 반도체 소자와 회로망의 품질검사, 고분자 및 유기물, 생체시료 nnnnnnnnn와 유가공 제품 등 모든 산업영역에 걸쳐 있다(Fig. 1). 입사된 전자빔이 시료의 원자와 탄성, 비탄성 충돌을 할 때 2차 전자(secondary electron)외에 후방산란전자(back scattered electron), X선, 음극형광 등이 발생하게 되는 이것을 통하여 topography (시료의 표면 형상), morphology(시료의 구성입자의 형상), composition(시료의 구성원소), crystallography (시료의 원자배열상태)등의 정보를 얻을 수 있다. SEM은 2차 전자를 이용하여 시료의 표면형상을 측정하고 그 외에는 SEM을 플랫폼으로 하여 EDS (Energy Dispersive X-ray Spectroscopy), WDS (Wave Dispersive X-ray Spectroscope), EPMA (Electron Probe X-ray Micro Analyzer), FIB (Focus Ion Beam), EBIC (Electron Beam Induced Current), EBSD (Electron Backscatter Diffraction), PBMS (Particle Beam Mass Spectrometer) 등의 많은 분석장치들이 SEM에 부가적으로 장착되어 다양한 시료의 측정이 이루어진다. 이 중 결정구조, 조성분석을 쉽고 효과적으로 할 수 있게 하는 X선 분석장치인 EDS를 SEM에 일체화시킨 장비와 EDS 및 PBMS를 SEM에 장착하여 반도체 공정 중 발생하는 나노입자의 형상, 성분, 크기분포를 측정하는 PCDS(Particle Characteristic Diagnosis System)에 대해 소개하고자 한다. - EDS와 통합된 SEM 시스템 기본적으로 SEM과 EDS는 상호보완적인 기능을 통하여 매우 밀접하게 사용되고 있으나 제조사와 기술적 근간의 차이로 인해 전혀 다른 방식으로 운영되고 있다. 일반적으로 SEM과 EDS는 별개의 시스템으로 스캔회로와 이미지 프로세싱 회로가 개별적으로 구현되어 있지만 로렌츠힘에 의해 발생하는 전자빔의 왜곡을 보정을 위해 EDS 시스템은 SEM 시스템과 연동되어 운영될 수 밖에 없다. 따라서, 각각의 시스템에서는 필요하지만 전체 시스템에서 보면 중복된 기능을 가지는 전자회로들이 존재하게 되고 이로 인해 SEM과 EDS에서 보는 시료의 이미지의 차이로 인한 측정오차가 발생한다(Fig. 2). EDS와 통합된 SEM 시스템은 중복된 기능인 스캔을 담당하는 scanning generation circuit과 이미지 프로세싱을 담당하는 FPGA circuit 및 응용프로그램을 SEM의 회로와 프로그램을 사용하게 함으로 SEM과 EDS가 보는 시료의 이미지가 정확히 일치함으로 이미지 캘리브레이션이 필요없고 측정오차가 제거된 EDS 측정이 가능하다. - PCDS 공정 중 발생하는 입자는 반도체 생산 수율에 가장 큰 영향을 끼치는 원인으로 파악되고 있으며, 생산수율을 저하시키는 원인 중 70% 가량이 이와 관련된 것으로 알려져 있다. 현재 반도체 공정 중이나 반도체 공정 장비에서 발생하는 입자는 제어가 되고 있지 않은 실정이며 대부분의 반도체 공정은 저압환경에서 이루어지기에 이 때 발생하는 입자를 제어하기 위해서는 저압환경에서 측정할 수 있는 측정시스템이 필요하다. 최근 국내에서는 CVD (Chemical Vapor Deposition) 시스템 내 파이프내벽에서의 오염입자 침착은 심각한 문제점으로 인식되고 있다(Fig. 3). PCDS (Particle Characteristic Diagnosis System)는 오염입자의 형상을 측정할 수 있는 SEM, 오염입자의 성분을 측정할 수 있는 EDS, 저압환경에서 기체에 포함된 입자를 빔 형태로 집속, 가속, 포화상태에 이르게 대전시켜 오염입자의 크기분포를 측정할 수 있는 PBMS가 일체화 되어 반도체 공정 중 발생하는 나노입자 대해 실시간으로 대처와 조치가 가능하게 한다.

• Journal of Dental Rehabilitation and Applied Science
• /
• v.28 no.4
• /
• pp.339-347
• /
• 2012

In previous studies, methods for enhancing cellular response on the Hydroxyapatite coated implant surface were described. In this study, the changes of surface characteristics such as surface roughness, contact angle, surface energy and surface morphology were observed when Hydroxyapatite coated Ti discs were immersed in NaCl solution for various time. Hydroxyapatite coated Ti discs were immersed in 0.9% NaCl solution for 7, 14 and 21 days at $37^{\circ}C$. The control group comprises dry identical discs not immersed in a solution. (n=3) All discs were dried in air completely and the surface roughness was measured using confocal laser scanning microscopy(CLSM). Static contact angle was recorded by video contact angle analyzer after dropping distilled water on the surface. The surface energy was calculated from contact angles of the three liquids. Surface was observed using a field emission-scanning electron microscope(FE-SEM). As a result, the surface roughness of immersed Hydroxyapatite coated Ti discs increased significantly and the contact angle decreased comparing with control group discs. The surface energy of immersed discs increased except for discs immersed for 14 days.

• Clinical and Experimental Reproductive Medicine
• /
• v.24 no.3
• /
• pp.301-309
• /
• 1997

The objective of this study was to determine the microtubule assembly and chromatin configuration during the first cell cycle in bovine oocytes following injection of spermatozoon, sperm head and tail. The microtubule and chromatin configuration was imaged with fluorescent labeled monoclonal ${\alpha}$-tubulin antibody and propidium iodide under laser scanning confocal microscope. Microtubule and chromatin dynamics in bovine oocytes following intracytoplasmic sperm injection (ICSI) were not different from those observed during in vitro fertilization (IVF). Following ICSI, the microtubular aster was observed around sperm midpiece. During pronuclear formation, the sperm aster was enlarged and seen around male and female pronuclei. At mitotic metaphase, the microtubular spindle assemble astral poles and chromosomes were aligned on the spindle equator. At mitosis, asters were concentrated to each spindle pole and they filled the cytoplasm. After injection of the isolated sperm head, the microtubular aster was not seen around sperm head in any cases (0/18). Instead, microtubules were organized from the cytoplasm, which filled the whole cytoplasm during pronuclear apposition. These microtubules seem to move male and female pronuclei. These results suggest that isolated sperm head can develop into normal pronucleus in mature bovine oocytes, and competent to participate syngamy with the ootid chromatin. The functional microtubules following isolated sperm head injection in bovine oocytes appeared to be organized solely from maternal stores.

• The Korean Journal of Mycology
• /
• v.38 no.1
• /
• pp.69-74
• /
• 2010

The 2 strains of Pleurotus ostreatus showing very severe and mild symptom of brown blotch were selected, and their F1 hybrid strains were made by mating of their single spore isolates. Hydrophobicity of fruiting body surface and brown blotch severity of the parental and their 11 F1 hybrid strains showing different level of severity of brown blotch were determined. Correlation coefficient (R) between disease severity and hydrophobicity were 0.68 and 0.70 on two independent experiments. Correlation coefficient between disease severity and disease incidence that was determined in the oyster mushroom farm were 0.46 and 0.52 on two independent experiments. When GFP-tagged Pseudomonas tolaasii was monitored with confocal microscope on cap surface of fruiting body, more cells of the pathogen were clustered on the severe strain than the mild strain, which indicates that the bacterial pathogen proliferates more on the severe strain. These results suggest that hydrophobicity of oyster mushroom fruiting body affects disease severity of the brown blotch disease; the longer the bacterial suspension stays on fruiting body surface more severe symptom of the blotch develops.

• The Korean Journal of Vision Science
• /
• v.20 no.4
• /
• pp.531-541
• /
• 2018

Purpose : To evaluate the acute cytotoxic effect of polyhexamethylene biguanide (PHMB), epigallocatechin gallate (EGCG) and PHMB/EGCG mixture on the cultured human corneal epithelial cell (HCEpiC). Methods : HCEpiCs were cultured in the media of HCEpiC containing 0.00001~0.005% PHMB, 0.001~5% EGCG and 0.00005% PHMB/0.05% EGCG mixture respectively for 30, 60, 120 and 240 min. Cultured HCEpiCs were fixed and stained with Draq5 and cell viability and apoptosis were evaluated using confocal microscope and ImageXpress $Ultra^{TM}$. Results : Cultured HCEpiC did not show cytotoxic effect at below 0.00005% PHMB and below 0.05% EGCG concentration. In the media containing 0.00005% PHMB/0.05% EGCG, acute cytototoxic effect was not found, whereas damaged HCEpiCS were increased and survival cells were decreased in the media incubated for 240 min. Conclusion : The mixture of 0.00005% PHMB/0.05% EGCG showed non acute cytotoxic effect on the cultured HCEpiCs, however it is needed to investigate its chronic cytotoxic effect.

• Restorative Dentistry and Endodontics
• /
• v.36 no.4
• /
• pp.290-299
• /
• 2011

Objectives: The usage of fluoride varnish for a moderate to low caries-risk group has not been well validated. This study aimed to evaluate the preventive and therapeutic efficacies of fluoride varnish on the initiated root caries. Materials and Methods: Ten premolars were sectioned into quarters, further divided into two windows, one of which was painted with Fluor Protector (1,000 ppm fluoride, Ivoclar Vivadent). An initial lesion with a well-preserved surface layer was produced by pH cycling. Scanned line analysis using energy dispersive spectrometry determined the weight percentages of Ca and P in the demineralized layer. Scanning Electron microscopy and confocal laser scanning microscopy (CLSM) evaluated the varnish-applied root surfaces. Results: The mean lesion depth (SD) was 12.3 (2.6) ${\mu}m$ (single cycling) and 19.6 (3.8) ${\mu}m$ (double cycling). Double cycling extended the lesion depth, but induced no more mineral loss than single cycling (p < 0.05). The mean weight percentages of Ca and P between groups with and without varnish were not significantly different (p < 0.05). A CLSM showed varnish remained within 15 ${\mu}m$ of the surface layer. Conclusions: When a mild acid challenge initiated root tissue demineralization, the application of low-concentration fluoride varnish did not influence the lesion depth or the mineral composition of the subsurface lesion.

• Korean Journal of Dental Materials
• /
• v.45 no.4
• /
• pp.257-274
• /
• 2018

The purpose of this study was to evaluate the effect of acid-treatment conditions on the surface properties of the RBM (Resorbable Blast Media) treated titanium. Disk typed cp-titanium specimens were prepared and RBM treatments was performed with calcium phosphate ceramic powder. Acid solution was mixed using HCl, $H_2SO_4$ and deionized water with 4 different volume fraction. The RBM treated titanium was acid treated with different acid solutions at 3 different temperatures and for 3 different periods. After acid-treatments, samples were cleaned with 1 % Solujet solution for 30 min and deionized water for 30 min using ultrasonic cleanser, then dried in the electrical oven ($37^{\circ}C$). Weight of samples before and after acid-treatment were measured using electric balance. Surface roughness was estimated using a confocal laser scanning microscopy, crystal phase in the surface of sample was analyzed using X-ray diffractometer. Surface morphology and components were evaluated using Scanning Electron Microscope (SEM) with Energy Dispersive X-ray spectroscopy (EDX) and X-ray Photoemission Spectroscopy (XPS). Values of the weight changes and surface roughness were statistically analyzed using Tukey-multiple comparison test (p=0.05). Weight change after acid treatments were significantly increased with increasing the concentration of $H_2SO_4$ and temperature of acid-solution. Acid-treatment conditions (concentration of $H_2SO_4$, temperature and time) did not produce consistent effects on the surface roughness, it showed the scattered results. From XRD analysis, formation of titanium hydrides in the titanium surface were observed in all specimens treated with acid-solutions. From XPS analysis, thin titanium oxide layer in the acid-treated specimens could be evaluated. Acid solution with $90^{\circ}C$ showed the strong effect on the titanium surface, it should be treated with caution to avoid the over-etching process.