• Journal of the Optical Society of Korea
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• v.18 no.5
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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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• 2000.11a
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• pp.191-193
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• 2000

A whole-cell biocatalyst was constructed by immobilizing an enzyme on the surface of the yeast Saccharomyces cerevisiae. The gene encoding Bacillus macerans cyclodextrin glucanotransferase(CGTase) was fused with the AGA2 gene encoding a small peptide disulfide-linked to the aga1, a cell wall protein of a-agglutinin. The plasmid was introduced S. cerevisiae and expressed in the medium consisting of 10g/L yeast extract, 20g/L peptone, and 20g/L galactose. The activity was detected with the formation of cyclodextrin(CD) from 10g/L soluble starch. Surface display of CGTase was also verified with the halo-test, flow cytometry, and immunofluorescence microscopy. The recombinant S. cerevisiae produced ${\alpha}-cyclodextrin$ more efficiently than the free CGTase by simultaneous fermentation and cyclization as yeast consumes glucose and maltose which are inhibitors for CD synthesis.

• Korean Journal of Microbiology
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• v.40 no.1
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• pp.23-28
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• 2004

Antigenic analysis of Herpes simplex type 2 virus was performed and its major antigen was localized using an immunoelectron microscopy. Antigens of 32, 43, 59 and 69 kDa were constantly expressed during the course of infection for 48 hr in the infected Vero cell. An antigen of 51 kDa was turned out to be the major one in inducing a immune response in Western-blot analysis. The 51 kDa antigen was localized on the surface of HSV-2 by immunoelectron microscopy using colloidal golds and anti-HSV 2 polyc1onal antibody. Immunofluorescence assay indicated that viral antigens were found throughout the infected cell and, especially, on the surface of the cell.

• Journal of Microbiology and Biotechnology
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• v.10 no.5
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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.

• Korean Journal of Veterinary Research
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• v.49 no.4
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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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• v.23 no.1
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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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• v.32 no.2
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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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• v.23 no.2
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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.

• The Korea Journal of Herbology
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• v.36 no.5
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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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• v.33 no.2
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• pp.105-116
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• 2003

In Vitro fertilization of U. unicinctus eggs observed by immunofluorescence and electron microscopes revealed an overview of the meiotic pattern of the tide animals. The eggs have been fertilized early at germinal vesicle stage, followed by germinal vesicle break down (GVBD), but pre-mitotic aster like structure could not be resolved by the methods employed in this work. The meiotic features, such as rotation-shift movement of spindle fibers, behavior of spermatozoonmonaster in the egg cytoplasm and active spindle fiber of the 1st polar body, have been observed. The antitubulin-FITC fluorescence show the 2nd meiotic apparatus appeared firstly parallel to the tangential line of the oolemma, proceeding the meiosis, its bipolarity is rotated and shifted towards the oolemma. The polar bodysite of the oolemma was not amorphous, but active in a sense of anti-tubulin-FITC reactions during the extrusions of the polar bodies. The immunofluorescence reactions of the spermatozoon centriole appeared at a later stage of the 2nd meiosis. During the time periods, the fertilized spermatozoon resided in the egg cytoplasm. Activating the centrioles, spermatozoon approaches towards the chromosomal materials of the 2nd oocyte. This suggests that spermatozoon centrioles initiate and play a roll to fuse male and female pronuclei.