• Applied Microscopy
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• v.38 no.3
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• pp.275-278
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• 2008

Electron tomography (ET) is an electron microscopic technique for obtaining a 3-D image from any electron microscopy specimen and its application in biomedical science has been increased thanks to development of electron microscopy and related technologies during the last decade. There are few researches on ET in Korea during this period. Although the importance of ET has been recognized recently by many researchers, initial approach to electron tomographic research is not easy for beginners. The 4th International Congress on Electron Tomography (4 ICET) was held on Nov $5{\sim}8$, 2006, at San Diego. The program dealt instrumentation, reconstruction algorithm, visualization/quantitative analysis and electron tomographic presentation of biological specimen and nano particles. 1 have summarized oral presentations and analyzed the posters presented on the meeting. Cryo-electron microscopic system was popular system for ET and followed conventional transmission electron microscopic system. Cultured cell line and tissue were most popular specimens analyzed and microorganisms including bacteria and virus also constituted important specimens. This analysis provides a current state of art in ET research and a guide line for practical application of ET and further research strategies.

• Proceedings of the Korea Information Processing Society Conference
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• 2009.11a
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• pp.395-396
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• 2009

Electron tomography를 이용한 3차원적 영상 시각화는 electron microscopy를 통해 하나의 실험 대상으로부터 연속된 이미지를 생산함으로써 이루어진다. 이미지 데이터 내부에는 대용량의 정보값을 포함하고 있어 3차원 구조물로의 변환이 가능하다. electron tomography 작업 과정 중 고해상도 원본 이미지에 pattern recognition 알고리즘이 적용된 필터링을 적용하면 실험에 필요한 데이터의 정보 손실을 최소화한 상태에서 electron tomography 시스템의 효율성을 높일 수 있다. 또한 tomographic econstruction이 진행되는 각 단계에 hanning windowing을 적용하면 불필요한 정보 값이나 노이즈 등을 효과적으로 제거할 수 있다. 윤곽선 데이터의 효과적 활용을 위하여 sobel 필터 처리를 할 경우 관찰하고자 하는 대상의 윤곽선 특징을 뚜렷하게 시각화 할 수 있었다. 본 연구를 통하여 데이터의 시각화 과정에서 실험의 신뢰성 확보를 위해 원본 이미지를 기반으로 하는 tomogram과 필터링을 적용한 tomogram을 비교하여 최종 결과물의 정확도를 높이고, electron tomography를 통한 결과물의 질적 향상을 유도할 수 있음을 확인하였다.

• Applied Microscopy
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• v.38 no.4
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• pp.285-291
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• 2008

We have evaluated the effects of experimental factors on transmitted electron beam intensities for quantitative analysis in electron tomography. For the correct application of Beer's law in electron tomography, the transmitted beam intensity should reflect the net effect of mass properties on beam path. So, the any other effects of the objective aperture and the specimen holder on beam path should be removed. The cut-off effects of objective aperture were examined using Quanti-foil holey carbon film and a transmission electron microscope operated at 120 kV. The transmitted beam intensities with $30{\mu}m$ objective aperture dropped about 16.7% compared to electron beam intensities without the objective aperture. Also, the additional losses of about 14.2% at high tilt angles were occurred by cut-off effects of the objective apertures. For the precise quantitative analysis in electron tomography, the effect of the objective aperture on transmitted electron beam intensities should be considered. It is desirable that 2-D tilt series images are obtained without the objective aperture for correct application of Bee's law.

• Applied Microscopy
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• v.42 no.4
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• pp.179-185
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• 2012

Alzheimer's disease (AD) is a progressive neurodegenerative disorder. Neuropathological hallmarks of AD are amyloid plaques, dystrophic neurite, and alteration of subcellular organelles. However, the morpho-functional study of this degenerative process and ultimate neuronal death remains poorly elucidated. In this study, immunohistochemical and ultrastructural analyses were performed to clarify the abnormal morphological alterations caused by the progression of AD in APP/PSEN1 transgenic mice, express human amyloid precursor protein, as a model for AD. In transgenic AD mice brain, the accumulation of Amyloid ${\beta}$ plaques and well-developed dystrophic neurites containing anti-LC3 antibody-positive autophagosomes were detected in the hippocampus and cortex regions. We also found severe disruption of mitochondrial cristae using high-voltage electron microscopy and three-dimensional electron tomography (3D tomography). These results provide morpho-functional evidence on the alteration of subcellular organelles in AD and may help in the investigation of the pathogenesis of AD.

• Applied Microscopy
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• v.50
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• pp.14.1-14.6
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• 2020

Scanning electron microscopy (SEM) plays a central role in analyzing structures by imaging a large area of brain tissue at nanometer scales. A vast amount of data in the large area are required to study structural changes of cellular organelles in a specific cell, such as neurons, astrocytes, oligodendrocytes, and microglia among brain tissue, at sufficient resolution. Array tomography is a useful method for large-area imaging, and the osmium-thiocarbohydrazide-osmium (OTO) and ferrocyanide-reduced osmium methods are commonly used to enhance membrane contrast. Because many samples prepared using the conventional technique without en bloc staining are considered inadequate for array tomography, we suggested an alternative technique using post-staining conventional samples and compared the advantages.

• Applied Microscopy
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• v.40 no.2
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• pp.117-123
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• 2010

This study has examined experimentally the Beer's law which is a precondition for quantitative electron tomography. We used carbon support film and latex spheres, which have similar absorption coefficients with biological samples, as the test samples to take a tilt-series of images for electron tomography. First, the 3D information of carbon film and latex spheres was obtained by electron tomography. Then, the regression analysis on the relationship between the intensities of the incident and the transmitted beams in a tilt series was carried out to examine the Beer's law. The regression results with RMS error of 0.976 show the linear intensity variations of the transmitted beam as the tilt angles were increased. In addition, the relative absorption coefficients of carbon support film and latex spheres calculated experimentally through the Beer's law were 1.71 (5) and 2.67 (6)/${\mu}m$, respectively. The absorption coefficients remained constant within a full tilt range. Therefore, it is expected that quantitative electron tomography could be performed for biological samples by applying Beer's law provided the exact intensity of incident beam can be obtained under the thoroughly controlled experimental conditions.

• Applied Microscopy
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• v.47 no.4
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• pp.223-225
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• 2017

Cryo-electron microscopy (cryo-EM) is arguably the most powerful tool used in structural biology. It is an important analytical technique that is used for gaining insight into the functional and molecular mechanisms of biomolecules involved in several physiological processes. Cryo-EM can be separated into the following three groups according to the analytical purposes and the features of the biological samples: cryo-electron tomography (cryo-ET), cryo-single-particle reconstruction, and cryo-electron crystallography. Cryo-tomography is a unique EM technique that is used to study intact biomolecular complexes within their original environments; it can provide mechanistic insights that are challenging for other EM-methods. However, the resolution of reconstructed three-dimensional (3D) models generated by cryo-ET is relatively low, while single-particle reconstruction can reproduce biomolecular structures having near-atomic resolution without the need for crystallization unless the samples are large (>200 kDa) and highly symmetrical. Cryo-electron crystallography is subdivided into the following two categories according to the types of samples: one category that deals with two-dimensional (2D) crystalline arrays and the other category that uses 3D crystals. These two categories of electron-crystallographic techniques use different diffraction data obtained from still diffraction and continuous-rotation diffraction. In this paper, we review crystal-based cryo-EM techniques and focus on the recently developed 3D electron-crystallographic technique called microcrystal-electron diffraction.

• 한국전자현미경학회:학술대회논문집
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• 2007.05a
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• pp.32-32
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• 2007

• Applied Microscopy
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• v.43 no.3
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• pp.122-126
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• 2013

The heterogeneous nucleation of the ${\Theta}^{\prime}$ phase on nanoscale precipitates has been investigated using a combination of three-dimensional atom probe tomography and high-resolution transmission electron microscopy. Two types of ${\Theta}^{\prime}$ phases were observed, namely small (~2 nm thick) cylindrical precipitates and larger (~100 nm) globular precipitates and both appear to be heterogeneously nucleated on the nanoscale precipitates. The composition and crystal structure of precipitates were directly analyzed by combination of two advanced characterization techniques.

• Applied Microscopy
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• v.43 no.1
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• pp.1-8
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• 2013

We developed an improved method for tilt series alignment with fiducial markers in electron tomography. Based on previous works regarding alignment, we adapted the Levenberg-Marquardt method to solve the nonlinear least squares problem by incorporating a new formula for the alignment model. We also suggested a new method to estimate the initial value for inversion with higher accuracy. The proposed approach was applied to geopolymers. A better alignment of the tilt series was achieved than that by IMOD S/W. The initial value estimation provided both stability and a good rate of convergence since the new method uses all marker positions, including those partly covering the tilt images.