• Nuclear Medicine and Molecular Imaging
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• v.41 no.3
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• pp.226-233
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• 2007

Purpose: Dual reporter gene imaging has several advantages for more sophisticated molecular imaging studies such as gene therapy monitoring. Herein, we have constructed hepatoma cell line expressing dual reporter genes of sodium iodide symporter (NIS) and enhanced green fluorescence protein (EGFP), and the functionalities of the genes were evaluated in vivo by nuclear and optical imaging. Materials and Methods: A pRetro-PN vector was constructed after separating NIS gene from pcDNA-NIS. RSV-EGFP-WPRE fragment separated from pLNRGW was cloned into pRetro-PN vector. The final vector expressing dual reporter genes was named pRetro-PNRGW. A human hepatoma (HepG2) cells were transfected by the retrovirus containing NIS and EGFP gene (HepG2-NE). Expression of NIS gene was confirmed by RT-PCR, radioiodine uptake and efflux studies. Expression of EGFP was confirmed by RT-PCR and fluorescence microscope. The HepG2 and HepG2-NE cells were implanted in shoulder and hindlimb of nude mice, then fluorescence image, gamma camera image and I-124 microPET image were undertaken. Results: The HepG2-NE cell was successfully constructed. RT-PCR showed NIS and EGFP mRNA expression. About 50% of cells showed fluorescence. The iodine uptake of NIS-expressed cells was about 9 times higher than control. In efflux study, $T_{1/2}$ of HepG2-NE cells was 9 min. HepG2-NE xenograft showed high signal-to-background fluorescent spots and higher iodine-uptake compared to those of HepG2 xenograft. Conclusion: A hepatoma cell line expressing NIS and EGFP dual reporter genes was successfully constructed and could be used as a potential either by therapeutic gene or imaging reporter gene.

• Childhood Kidney Diseases
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• v.15 no.1
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• pp.29-37
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• 2011

To understand the course of renal diseases well, we must have basic knowledges of histologic procedures of renal biopsy samples as well as basic pathologic changes. This article describes the method of dividing the biopsy samples, fixatives for various pathologic examinations and basic pathologic changes of glomerular diseases. For light microscopic examination, color changes of glomerular structures in PAS, trichrome and PAM stains, normal glomerular patterns compared to various glomerulopathies are introduced. While describing typical staining patterns and intensities of fluorescence in membranous glomerulopathy and IgA nephropathy, basic interpretation of immunofluorescent microscopic examination is described. To understand electron microscopic pictures of renal diseases, preference locations of electron dense deposits in various glomerulonephrites are described with schema. This article is the introduction part of the renal pathology and for the further detail changes of specific entities, we should reference the renal pathology textbooks or articles.

• Journal of the korean academy of Pediatric Dentistry
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• v.30 no.2
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• pp.189-195
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• 2003

The objective of this study is to improve the optical sensitivity of laser fluorescence for detection of incipient enamel caries. An incipient carious lesion was formed in various stages by placing an enamel specimen of a bovine tooth in STPP demineralization solution. After measuring the optical density of the lesion surface by laser fluorescence induced by argon laser and various alter of yellow(500-520nm), amber(520-540nm), orange(540-560nm), and red(560-580nm), the specimen was cut vertically to measure the depth of the lesion using a polarizing microscope. SAS statistical program was used to analyze the relationship between the optical density of the lesion suface and the depth of the lesion. The results were as follows: 1. The optical density of early carious lesion, measured by laser fluorescence with amber and orange filter, and lesion depth observed by polarizing microscope, were increased as demineralization time increased. 2. The correlation coefficient between optical density of the lesion surface and the histological depth of the lesion was the highest in orange filter(r=0.49), followed by amber(r=0.32), yellow(r=0.13) and red(0.01). 3. Regression analysis showed that the most linear relationship between the optical density and the lesion depth was existed in orange filter group. In regard above results, laser fluorescence could be considered to be reliable for optical diagnosis of dental caries.

• Journal of the Korea Computer Graphics Society
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• v.23 no.4
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• pp.31-40
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• 2017

As 3D cell culture has recently become possible, it has been able to observe a 3D shape of cell and volume. Generally, 3D information of a cell should be observed with a special microscope such as a confocal microscope or an electron microscope. However, a confocal microscope is more expensive than a conventional microscope and takes longer time to capture images. Therefore, there is a need for a method that can reconstruct the 3D shape of cells using a common microscope. In this paper, we propose a method of reconstructing 3D cells using the focus estimator value from multi-focal fluorescence images of cells. Initially, 3D cultured cells are captured with an optical microscope by changing the focus. Then the approximate position of the cells is assigned as ROI (Region Of Interest) using the circular Hough transform in the images. The MSBF (Modified Sliding Band Filter) is applied to the obtained ROI to extract the outlines of the cell clusters, and the focus estimator values are computed based on the extracted outlines. Using the computed focus estimator values and the numerical aperture (NA) of the microscope, we extract the outline of the cell cluster considering the depth and reconstruct the cells into 3D based on the extracted outline. The reconstruction results are examined by comparing with the combined in-focus portions of the cell images.

• Proceedings of the Korea Water Resources Association Conference
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• 2022.05a
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• pp.261-261
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• 2022

최근 국내 녹조현상의 빈도가 잦아지고, 지속기간이 증가하였다. 이러한 녹조현상은 이상기후로 인한 일사량, 수온, 체류시간 증가로 인한 환경적인 영향이 있으며, 산업단지 및 농지 등에서 유출되는 질소나 인의 부영양화로 인한 인위적 영향이 있다. 조류종 중 특히 남조류가 생장하는 과정에서 발생하는 독성물질(간독소, 신경독소), 이취미 물질(Geosmin, 2-MIB 등)은 수질을 악화시키고 있어 생태계에 큰 영향을 미친다. 조류는 하천에서 넓은 분포로 발생하게 되는데 현재 조류 조사 방법은 현장에 발생한 조류를 직접 채취하여 연구실에서 현미경으로 직접 검경하기 때문에 많은 인력과 시간, 비용이 소비되고 있는 실정이다. 국내에서는 인력과 시간을 줄이기 위해 센서를 활용한 조류분석 및 원격탐사 기법을 이용한 조류 모니터링에 대한 연구가 많이 진행되고 있다. 하지만 하천에서는 녹조류와 남조류 모두 혼합되어 존재하며 센서를 이용하여 조류를 분석할 시 녹조류는 Chl-a, 남조류는 Phycocyanin을 대체하여 분석하고 있다. 하지만 모든 조류는 광합성을 하기 때문에 Chl-a를 모두 함유하고 있어 Chl-a만으로 남조류를 추정하는 데에는 한계가 있다. 따라서 본 연구에서는 조류 계측장비인 LISST-HAB를 통하여 조류시료의 농도를 측정하여 Chl-a, Phycocyanin 농도를 측정하여 조류종 분류에 대한 분석을 진행하였다.

• Polymer(Korea)
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• v.29 no.6
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• pp.543-548
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• 2005

Double-layered spheres play an important role in controlling drug delivery for pharmaceutical application, because of the low initial burst compared with single-layered spheres and targetable delivery to specific organ. But it has drawback in loading drug and controlling size. In this study, we developed double-layered spheres using relatively simple oil-in-water (O/W) solvent evaporation method witw/without ultrasonication and investigated the size variation of the double-layered microspheres on the contents of poly(lactide- co-glycolide) (PLGA). Double - layered spheres were char-acterized by scanning elecron microscope (SEM), camscope, and confocal fluorescence laser microscope (CFLM). Double-layered spheres showed smooth surfaces and obvious difference between core and corona by SEM observation and camscope. We observed the fluorescent core in the double-walled spheres composed of FlTC-dextran and PLGA using CFLM. It was found that the core of the microsphere was dextran and the corona of the fabricate microsphere was PLGA. Also, the more PLGA concentration, the more the size of the fabricating double-layered sphere observed.

• Journal of the korean academy of Pediatric Dentistry
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• v.46 no.3
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• pp.265-273
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• 2019

This study was performed to evaluate clinical use of laser fluorescence (LF) to identify early childhood caries lesions suitable for applying resin infiltration. 20 exfoliated primary molars with proximal caries were selected and cut buccolingually cross the central pit for regarding the mesial and distal surfaces respectively. 27 specimens corresponding to ICDAS code 1 and 2 were selected and the LF values were measured. When infiltrant resin was applied, double staining for microscopy detection has done simultaneously. Tooth samples were sliced with 0.7 mm thick. The maximum lesion depth, maximum penetration depth, and average penetration rate were measured from the confocal scanning laser microscope image. Pearson correlation analysis was performed. The intraclass correlation coefficient of LF values shows excellent agreement. LF values had positive correlation with penetration rate, but not lesion depth and penetration depth. Significant correlation between LF readings and penetration rate was verified in deep enamel caries and dentin caries except shallow enamel caries. Infiltrant resin could penetrate with a higher rate and LF values could be increased in more active caries lesions. In assessing radiologically similar caries lesion, laser fluorescence might be useful for identifying caries activity.

• Journal of the Microelectronics and Packaging Society
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• v.20 no.2
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• pp.23-28
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• 2013

Microlens array as basic element of the optical system have been fabricated with various focal length (mainly with long focal length) depending on the purpose of application. In this paper, the microlens arrays were fabricated for observing fluorescent images within sol-gel. Though the fluorescent signal is very low, the microlens array can help obtaining clear images through extracting the fluorescent light from sol-gel. We fabricated microlens arrays with short focal length, which can extract the light using photoresist thermal reflow method. In the experiment, the diameter of microlens decreased after thermal reflow because the solvent within the photoresist was vaporized. Therefore, to compensate the shape error by this reduction, microlens diameter in photomask was altered and spin-coat recipe of photoresist were modified.

• Proceedings of the Korean Vacuum Society Conference
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• 2014.02a
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• pp.422.2-422.2
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• 2014

가스장 이온원(GFIS: Gas Field Ionization Source)은 전자현미경보다 분해능이 향상된 이온현미경의 광원으로 사용하기 위하여 연구되고 있고, 큰 각전류 밀도, 작은 크기의 가상 이온원 그리고 좁은 에너지 퍼짐을 특징으로 한다. 여러 가지 장점을 가지고 있는 GFIS을 개발하기 위해서는 GFIS에서 발생된 이온빔의 형상을 관찰 것이 매우 중요하며, 이러한 관찰을 위한 시스템에는 주로 마이크로 채널 플레이트 (MCP: Micro Channel Plate)가 사용된다. MCP는 채널내부에 입사한 입자의 에너지에 의해서 생성된 이차전자를 수 천 배에서 수 백 만 배 이상 증폭시켜 형광판에 조사하고 발광시키는 방법으로 작은 신호를 영상으로 관찰 할 수 있도록 한다. MCP의 큰 증폭비는 작은 크기의 신호를 큰 신호로 증폭하여 관찰하는데 용이하여, GFIS 방법으로 생성된 이온빔(이온빔 전류 값은 pA 수준)을 관찰하기에 적합하다. 그러나 MCP를 이용하여도 증폭된 이온빔의 세기가 매우 작기때문에 생성된 이온빔 형상을 정확하게 관찰하기 위해서는 MCP의 형광판을 촬영하는 카메라 노출시간을 길게하여 데이터 수집 시간을 늘려야 하는 문제가 있다. 본 발표에서는 이온빔 형상 관찰에 소요되는 시간을 단축하기 위하여 MCP의 잡음이 GFIS의 이온빔 이미지 관찰에 미치는 영향을 분석하고 이를 제거 방법을 소개한다. 본 연구에서는 GFIS 방출 이온빔의 이미지에 포함된 MCP 잡음 특성을 장(전계)이온현미경 (Field Ion Microscope)실험을 통하여 분석하였고, 디지털 이미지 처리 방법을 이용하여 방출 이온빔 이미지에서 MCP 잡음을 제거하여 방출 이온빔 이미지만 추출할 수 있었다. 본 연구에서 제안한 방법을 GFIS 방출 이온빔 관찰시스템에 적용함으로써 기존 방법에 비해 노출시간을 단축하여 방출 이온빔을 관찰 할 수 있었으며, 노이즈 제거 효과로 향상된 이온빔 형상을 얻을 수 있었다. 본 연구결과의 관찰시간 단축과 향상된 이온빔 형상 획득은 이온현미경 개발에 필수적인 단원자 이온빔을 보다 효율적으로 개발할 수 있으며 디지털 이미지 처리로 GFIS 이온빔 생성을 자동화하는데 응용할 수 있다. 더불어 기존방법에 비해 이미지 획득을 위한 MCP의 노출시간을 단축할 수 있으므로 실험장비 수명 단축 방지 및 관리에 큰 장점이 있다.

• Journal of the Korean Society of Visualization
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• v.12 no.3
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• pp.9-14
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• 2014

Two-photon microscopy (TPM) has been used in plant research as a high-resolution high-depth 3D imaging modality. However, TPM is known to induce photo-damage to the plant in case of long time exposure, and optimal excitation wavelength for plant imaging has not been investigated. Longer excitation wavelength may be appropriate for in vivo two-photon imaging of Arabidopsis thaliana leaves, and effects of longer excitation wavelength were investigated in terms of imaging depth, emission spectrum. Changes of emission spectrum as a function of exposure time at longer excitation wavelength were measured for in vivo longitudinal imaging. Imaging depth was not changed much probably because photon scattering at the cell wall was a limiting factor. Chloroplast emission spectrum showed its intensity peak shift by 20 nm with transition of excitation wavelength from 849 nm or below to 850 nm or higher. Emission spectrum showed different change patterns with excitation wavelengths in longitudinal imaging. Longer excitation wavelengths appeared to interact with chloroplasts differently in comparison with 780 nm excitation wavelength, and may be good for in vivo imaging.