• Journal of the Optical Society of Korea
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• 제18권5호
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• pp.551-557
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• 2014

In this study, we demonstrated multimodal nonlinear optical (NLO) microscopy integrated simultaneously with two-photon excitation fluorescence (TPEF), second-harmonic generation (SHG), and coherent anti-Stokes Raman scattering (CARS) in order to obtain targeted cellular and label-free images in an immunofluorescence assay of the atherosclerotic aorta from apolipoprotein E-deficient mice. The multimodal NLO microscope used two laser systems: picosecond (ps) and femtosecond (fs) pulsed lasers. A pair of ps-pulsed lights served for CARS (817 nm and 1064 nm) and SHG (817 nm) images; light from the fs-pulsed laser with the center wavelength of 720 nm was incident into the sample to obtain autofluorescence and targeted molecular TPEF images for high efficiency of fluorescence intensity without cross-talk. For multicolor-targeted TPEF imaging, we stained smooth-muscle cells and macrophages with fluorescent dyes (Alexa Fluor 350 and Alexa Fluor 594) for an immunofluorescence assay. Each depth-sectioned image consisted of $512{\times}512$ pixels with a field of view of $250{\times}250{\mu}m^2$, a lateral resolution of $0.4{\mu}m$, and an axial resolution of $1.3{\mu}m$. We obtained composite multicolor images with conventional label-free NLO images and targeted TPEF images in atherosclerotic-plaque samples. Multicolor 3-D imaging of atherosclerotic-plaque structural and functional composition will be helpful for understanding the pathogenesis of cardiovascular disease.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.191-193
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• 2000

B. macerans 유래의 CGTase를 yeast surface display기술을 이용하여 S. cerevisiae의 표면에 발현된 것을 halo-test와 immunofluorescence microscopy와 flow cytometry를 통하여 확인하였다. 재조합 효모는 효소의 cyclization작용을 저해하고 CD의 분해작용을 촉진하는 glucose와 maltose를 제거하는 발효공정과 표면 발현된 CGTase의 cyclization 공정을 동시에 수행할 수 있어 CD의 생산, 분리공정을 효율적으로 개선하였다.

• 미생물학회지
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• 제40권1호
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• pp.23-28
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• 2004

단순포진 바이러스 감염을 유발하는Herpes simplex2형 바이러스의 감염에 관여하는 항원들과 중화항체 생산을 유발하는 주요 항원들의 위치를 확인하였다. Vero cell에 감염하였을 때 48시간 동안 31, 43, 59, 69 kDa 바이러스 항원들이 지속적으로 발현되었으며, 감염된 쥐에서 생산한 항체와의 반응에서는 51 kDa 항원이 가장 강한 반응을 보였다. 면역전자현미경으로 위치를 확인한 결과 colloidal gold가 바이러스 표면에 발견되는 것으로 보아 이 항원이 바이러스 표면에 존재하고 있는 것으로 확인되었다. 형광현미경 분석은 이 항원들이 감염된 세포 내에서 전반적으로 발견되었고 특히 세포 표면에서 많이 발현되고 있었다.

• Journal of Microbiology and Biotechnology
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• 제10권5호
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• pp.714-720
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• 2000

Antigens of Herpes simplex virus type 1 (HSV-1) strain F were immunoblotted to identify the most immunodominant one, and the localization of this antigen was then studied using immunoelectron microscopy. The 67.8 kDa antigen appeared to be the most immunodominant one in a mouse model, and it showed randomly scattered and partially clustered distribution on the surface of the virion. The localization study was performed using immunogold with polyclonal anti-HSV-1 sera produced from BALB/c mice, and immunofluorescence demonstrated that the viral products in the HSV-2 infected Vero cells were distributed throughout the infected host cell, however, mainly on the surface of the host membrane.

• 대한수의학회지
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• 제49권4호
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• pp.273-278
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• 2009

Neurofascin, one of the members of L1CAM, has been known to have some important roles during the development of nerve fibers. In order to investigate the role of neurofascin associated with the formation of Schmidt-Lantermann incisure in the sciatic nerve, the localization of neurofascin was studied with electron microscopy, immuno-fluorescence and immuno-electron microscopy. In the electron microscopy, the first formation of Schmidt-Lantermann incisure was checked at postnatal day 6 and the complete form of incisures traversing the whole myelin sheath began to be observed at postnatal day 8. In the immunofluorescence, neurofascin immunoreactive Schmidt-Lantermann incisures were first checked at postnatal day 6 and dramatically increased with aging by postnatal day 56. In the immunoelectron microscopy, neurofascin immunoreactive gold particles at the incisure forming sites were first observed at postnatal day 6 and the number of gold particles was increased as the animal was getting old by postnatal day 56. According to the present study, neurofascin is likely to have some relationships with Schmidt-Lantermann incisure formation.

• Applied Microscopy
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• 제23권1호
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• pp.25-34
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• 1993

Prolactin has been reported to be present in the tear film of humans and prolactin-like immunoreactivity has been detected by immunofluorescence in acinar cells of the lacrimal glands of humans and rats. The present study was aimed at clarifying the intracellular distribution of the prolactin-like immunoreactivity, using the electron microscope immunogold technique. The lacrimal gland acinar cells have two types of secretory granules: 1) Secretory granules containing flocculent materials irregularly shaped and are often coalesced. 2) Secretory granules are fairly round and contain homogenous materials of a moderate electron density. The density of the granular content varies even within a single cell. We found prolactin-like reactivity in secretory granules, some smaller cytosolic vesicles, Golgi cisternae and nuclei in acinar cells from intact glands of rat. Our present results are consistent with the conclusion that prolactin is present in lacrimal cells. The presence of prolactin reactivity in the nucleus suggests that prolactin may be a regulatory factor modulating gene expression.

• Journal of Trauma and Injury
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• 제32권2호
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• pp.111-114
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• 2019

Minimal change disease (MCD) in children has a favorable long-term prognosis, and development of end-stage renal disease is very uncommon; less than 5%. In the first case of its kind, we report a 21-year-old female with a history of MCD at the age of 6, who had late relapse subsequent to a motorcycle accident resulting in a de-gloving skin injury and intensive care unit admission. MCD was confirmed by normal light microscopy, podocyte effacement on electron microscopy and absence of any deposits on immunofluorescence 3 weeks after the incident due to critical illness. It is postulated that the skin injury is what caused the relapse of MCD.

• Childhood Kidney Diseases
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• 제23권2호
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• pp.93-99
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• 2019

C3 glomerulopathy is a renal disorder involving dysregulation of alternative pathway complement activation. In most instances, a membranoproliferative pattern of glomerular injury with a prevalence of C3 deposition is observed by immunofluorescence microscopy. Dense deposit disease (DDD) and C3 glomerulonephritis (C3GN) are subclasses of C3 glomerulopathy that are distinguishable by electron microscopy. Highly electron-dense transformation of glomerular basement membrane is characteristic of DDD. C3GN should be differentiated from post-infectious glomerulonephritis and other immune complex-mediated glomerulonephritides showing C3 deposits.

• 대한본초학회지
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• 제36권5호
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• pp.59-67
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• 2021

Objectives : Hibiscus (Hibiscus sabdariffa L.) is rich in antioxidants such as flavonoids and anthocyanins and is known to have anti-inflammatory activity and anti-aging function of the skin, but there is no study on its effect on the skin barrier. This study aim to investigate the positive effect on the skin barrier by confirming the effect of water extracts of Hibiscus sabdariffa L. (WEHS) on the tight junction-related gene expression. Methods : The antioxidant efficacy of WEHS was investigated through ABTS and DPPH assays, and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium was performed to examine the effect on cell viability. quantitative Reverse transcription polymerase chain reaction and immunoblot analysis were performed to confirm the effect of WEHS on the expression of tight junction-related genes, and immunofluorescence microscopy was used to confirm the movement of Claudin 1 protein into tight junctions. Results : WEHS showed strong antioxidant activity and induced an increase in both mRNA and protein expression levels of Claudin 1 among tight junction-related genes. The strong localization of Claudine 1 protein increased by WEHS to the tight junction was confirmed by immunofluorescence microscopy. Conclusions : Hibiscus was confirmed through this study to show antioxidant activity and the function of promoting the expression of the tight junction Claudin 1 gene, suggesting that Hibiscus can be used as a material for the prevention and treatment of skin moisturizing and atopy, which have an important influence on tight junction.

• Applied Microscopy
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• 제33권2호
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• pp.105-116
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• 2003

U. unicinctus 난세포를 인공수정한 결과 수정 시기는 germinal vesicle (2n)기였으며 정자 중앙부에서 돌출한 actomere와 난모세포의 미세융모 끝에서 세포막 융합이 시작되는 것으로 관찰되었다. Germinal vesicle기에 수정이 가능하였으므로 pre-mitotic spindle이 관찰되지 않은 것으로 사료되었다. 수정 후 난모세포는 제1, 2감수분열을 수행하였으며 각각의 감수분열 장치들은 감수분열 말기에 난모세포막과 밀접한 구조를 형성하였고, 이 부위에서 극체가 형성되는 것을 볼 수 있었다. 극체 형성시 난모세포막은 세포분열의 관점에서 형태가 없는 것이 아니라 항-튜블린-FITC에 대한 활성구조를 형성하였으며, 각 감수분열 장치의 한쪽 극 (pole)과 어떤 복합구조를 형성하는 것으로 보인다. 제2극체 형성도 제1극체와 유사한 방법으로 형성되었으나, 제2감수분열 장치는 난모세포막의 접선에 평행하게 생성된 후 세포막을 향해 이동하면서 방추사의 양극성이 회전 (shift-rotation)하였고 접선에 수직으로 정렬하는 것을 볼 수 있었다. 난모세포의 감수분열 장치에서 방추사의 활성은 강하였으나 aster의 활성은 비교적 약한 것으로 보였다. 제2감수분열이 진행되는 동안 난모세포질에 머믈고 있던 정자는 점차 미래의 자성 전핵 형성부인 난모세포막 근접부로 이동하는 것이 관찰되었다. 난모세포막 근접부로 이동하는 동안 정자 aster의 활성은 점차 강해지는 반면 난모세포의 aster에서는 활성이 미약한 것으로 보아서 자웅 전핵융합을 주도하는 것은 정자 유래의 aster인 것으로 사료되었다. 제1난모세포의 감수분열 장치가 활성화되는 시기에 제1극체의 방추사에서도 강한 활성을 관찰할 수 있었다.