• Journal of the Korean Society of Visualization
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• v.21 no.3
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• pp.65-74
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• 2023

Compared to epifluorescence(EPI) microscopy which captures fluorescence from the entire depth of sample, total internal reflection fluorescence(TIRF) can selectively visualize only a single surface of it. TIRF uses a thin evanescent field generated by the total internal reflection of laser light on surface. However, conventional TIRF system are designed for total internal reflection to occur at the upper surface of sample, making them unsuitable for sessile droplet imaging. We designed a TIRF system suitable for a sessile droplet imaging by utilizing slide glass as a lightguide. We presented the details for constructing the TIRF system using a prism, slide glass, air slit, and optical trap. Then, we compared the TIRF with EPI by imaging the droplet with fluorescent particles during its drying process. As a result, TIRF allows us to distinctly visualize the drying pattern on the bottom surface of droplet.

• Journal of the Optical Society of Korea
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• v.10 no.4
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• pp.178-183
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• 2006

An in-line phase-shifting digital holographic microscopy system was constructed by inserting a conventional microscope in the object arm of a Mach-Zehnder interferometer. It was used to measure the three dimensional shape of a micro Fresnel lens. It was also shown that both the lateral and the axial resolutions of the in-line phase-shifting system using a self-calibration algorithm were superior to those of the best off-axis system.

• Applied Microscopy
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• v.44 no.1
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• pp.8-14
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• 2014

The auditory system of the Korean greater horseshoe bat (Rhinolophus ferrumequinum korai, RFK) is adapted to its own echolocation signal, which consist of constant frequency (CF) element and frequency modulated (FM) element. In contrast, the Japanese long-fingered bat (Miniopterus schreibersii fuliginosus, MSF) emits FM signals. In the present study, the characteristics of stereocilia in RFK (a CF/FM bat) and MSF (a FM bat) were studied in the apical turn of the cochlea where the lower frequencies are transduced. Stereocilia lengths and numbers were quantitatively measured in RFK by scanning electron microscopy and compared with those of MSF. Each inner hair cells (IHCs) of RFK possessed three rows of stereocilia, whereas MSF possessed five rows of stereocilia. Gradients in stereocilia lengths and numbers of stereocilia of the IHCs of RFK were found to be less pronounced and fewer, respectively, than those of MSF. Each outer hair cells (OHCs) possessed three rows of stereocilia in both species. OHCs stereocilia in RFK that distinguished it from MSF were a shorter length and a greater number of stereocilia. These features suggest that the apical cochleas of RFK are adapted for the processing of higher frequency echolocation calls rather than that of MSF.

• Proceedings of the Korea Information Processing Society Conference
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• 2008.11a
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• pp.100-103
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• 2008

한국기초과학지원연구원(KBSI, Korea Basic Science Institute)에서는 국내 유일의 초고전압투과전자현미경(HVEM, High Voltage Electron Microscopy)을 비롯하여 3대의 일반투과 전자현미경을 보유하고 있다. 전자현미경을 통하여 관찰된 이미지는 각 단계별로 tilting 되어 저장된 이미지로서 관찰자에게 보다 나은 관찰 환경의 구성을 위해 3D로의 reconstruction은 필수 과정이라고 할 수 있겠다. 이 과정 중 카메라 중심에서 벋어난 부분의 왜곡을 워핑기법을 통하여 최대한 감소시킨다. 이런 전처리 과정을 통하여 3D 구조물을 구성하게 되면 초기 이미지를 그대로 사용하는 것보다 한 단계 더 나은 결과물을 얻어낼 수 있다. 이미지 전처리를 이용한 전자현미경 볼륨 랜더링 시스템의 구축은 관찰자에게 보다 편리하며 빠른 실험 환경을 제공하여 줄 수 있고, 이해하기 쉽고 실제 모습에 가까운 형태의 실험 결과물을 접할 수 있게 된다.

• Proceedings of the Korean Information Science Society Conference
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• 2005.07a
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• pp.583-585
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• 2005

바이오 및 재료 분야 등 NT(Nano Technology), BT(Bio Technology)에 관련된 연구를 보다 더 수월하게 하기 위해 본원에 설치되어 있는 초고전압 투과전자현미경(High Voltage Electron Microscopy - 이하 HVEM)을 이용한다. HVEM을 통해 획득한 이미지의 정보는 매우 방대하여 하나의 시료를 관찰하는데 있어서 수백 메가 이상의 용량을 차지하고, 연구를 수행하는데 있어서 데이터를 여러 형태로 관찰 분석하기 때문에 수월한 지원을 위해 체계적으로 관리할 수 있는 데이터베이스 시스템이 필요하다. 그러나 일반적인 범용 데이터베이스로는 이러한 대규모의 데이터를 저장할 수 없다. 따라서 본 논문에서는 이러한 용량 데이터를 체계적으로 관리할 수 있도록, 데이터 그리드와 연구 데이터의 정보를 갖는 metadata 테이블을 통해 서로 먼 거리에 있는 연구원들이 데이터를 접근하고 대규모 저장 공간을 갖는 데이터베이스 시스템을 제안한다.

• Applied Microscopy
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• v.48 no.4
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• pp.81-86
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• 2018

The respiratory chain complex forms a supercomplex (SC) in the inner mitochondrial membrane. This complex facilitates the process of electron transfer to produce the proton gradient used to synthesize ATP. Understanding the precise structure of the SC is considered an important challenge. However, it has not yet been reported. The development of a Cryo-electron microscopy (EM) technique provides an effective way to obtain high-resolution micrographs to determine the high-resolution three-dimensional structure of biomolecules. In this brief review, the currently reported Cryo-EM structures of the mammalian respirasome have been described in order to establish a direction for further research in the respiratory system.

• Applied Microscopy
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• v.52
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• pp.4.1-4.11
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• 2022

Field emission scanning electron microscopy (FESEM) is an essential tool for observing surface details of specimens in a high vacuum. A series of specimen procedures precludes the observations of living organisms, resulting in artifacts. To overcome these problems, Takahiko Hariyama and his colleagues proposed the concept of the "nanosuit" later referred to as "NanoSuit", describing a thin polymer layer placed on organisms to protect them in a high vacuum in 2013. The NanoSuit is formed rapidly by (i) electron beam irradiation, (ii) plasma irradiation, (iii) Tween 20 solution immersion, and (iv) surface shield enhancer (SSE) solution immersion. Without chemical fixation and metal coating, the NanoSuit-formed specimens allowed structural preservation and accurate element detection of insulating, wet specimens at high spatial resolution. NanoSuit-formed larvae were able to resume normal growth following FESEM observation. The method has been employed to observe unfixed and uncoated bacteria, multicellular organisms, and paraffin sections. These results suggest that the NanoSuit can be applied to prolong life in vacuo and overcome the limit of dead imaging of electron microscopy.

• Applied Microscopy
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• v.51
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• pp.16.1-16.7
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• 2021

Electron microscopy (EM) is an essential imaging method in biological sciences. Since biological specimens are exposed to radiation and vacuum conditions during EM observations, they die due to chemical bond breakage and desiccation. However, some organisms belonging to the taxa of bacteria, fungi, plants, and animals (including beetles, ticks, and tardigrades) have been reported to survive hostile scanning EM (SEM) conditions since the onset of EM. The surviving organisms were observed (i) without chemical fixation, (ii) after mounting to a precooled cold stage, (iii) using cryo-SEM, or (iv) after coating with a thin polymer layer, respectively. Combined use of these techniques may provide a better condition for preservation and live imaging of multicellular organisms for a long time beyond live-cell EM.

• Proceedings of the Korean Society of Precision Engineering Conference
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• 2005.10a
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• pp.576-580
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• 2005

New real-time confocal microscopy using spectral encoding technique and slit confocal aperture is proposed and designed. Spectral encoding technique, which encodes one-dimensional spatial information of a specimen in wavelength, and slit aperture make it possible to obtain two-dimensional lateral image of the specimen simultaneously at standard video rates without expensive scanning units such as polygon mirrors and galvano mirrors. The working principle and the configuration of the system are explained. The variation in axial responses for the simplified model of the system with normalized slit width is numerically analyzed based on the wave optics theory. Slit width that directly affects the depth discrimination of the system is determined by a compromise between axial resolution and signal intensity from the simulation result. On the assumption of the lateral sampling resolution of 50 nm, design variables and governing equations of the system are derived. The system is designed to have the mapping error less than the half pixel size, to be diffraction-limited and to have the maximum illumination efficiency. The designed system has the FOV of $12.8um{\times}9.6um$, the theoretical axial FWHM of 1.1 um and the lateral magnification of-367.8.

• Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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• 1998.11a
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• pp.133-136
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• 1998

The scanning Maxwell-stress microscopy (SMM) is a dynamic noncontact electric force microscopy that allows simultaneous access to the electrical properties of molecular system such as surface potential, surface charge, dielectric constant and conductivity along with the topography. Here we report our recent results of its application to nanoscopic study of domain structures and electrical functionality in organic thin films prepared by the Langmuir-Blodgett technique.