• 제목/요약/키워드: serial sectioning

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Sentinel Node Biopsy Examination for Breast Cancer in a Routine Laboratory Practice: Results of a Pilot Study

  • Khoo, Joon-Joon;Ng, Chen-Siew;Sabaratnam, Subathra;Arulanantham, Sarojah
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권3호
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    • pp.1149-1155
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    • 2016
  • Background: Examination of sentinel lymph node (SLN) biopsies provides accurate nodal staging for breast cancer and plays a key role in patient management. Procurement of SLNs and the methods used to process specimens are equally important. Increasing the level of detail in histopathological examination of SLNs increases detection of metastatic tumours but will also increase the burden of busy laboratories and thus may not be carried out routinely. Recommendation of a reasonable standard in SLN examination is required to ensure high sensitivity of results while maintaining a manageable practice workload. Materials and Methods: Twenty-four patients with clinically node-negative breast cancer were recruited. Combined radiotracer and blue dye methods were used for identification of SLNs. The nodes were thinly sliced and embedded. Serial sectioning and immunohistochemical (IHC) staining against AE1/AE3 were performed if initial H&E sections of the blocks were negative. Results: SLNs were successfully identified in all patients. Ten cases had nodal metastases with 7 detected in SLNs and 3 detected only in axillary nodes (false negative rate, FNR=30%). Some 5 out of 7 metastatic lesions in the SLNs (71.4%) were detected in initial sections of the thinly sliced tissue. Serial sectioning detected the remaining two cases with either micrometastases or isolated tumour cells (ITC). Conclusions: Thin slicing of tissue to 3-5mm thickness and serial sectioning improved the detection of micro and macro-metastases but the additional burden of serial sectioning gave low yield of micrometastases or ITC and may not be cost effective. IHC validation did not further increase sensitivity of detection. Therefore its use should only be limited to confirmation of suspicious lesions. False negative cases where SLNs were not involved could be due to skipped metastases to non-sentinel nodes or poor technique during procurement, resulting in missed detection of actual SLNs.

Structural Analysis of Exosomes Using Different Types of Electron Microscopy

  • Choi, Hyosun;Mun, Ji Young
    • Applied Microscopy
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    • 제47권3호
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    • pp.171-175
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    • 2017
  • Negative staining has been traditionally used for exosome imaging; however, the technique is limited to surface topology only and can cause staining artifacts. Therefore, to analyze the internal structure of exosomes, we employed a method of block preparation, thin sectioning, and electron tomography. In addition, an automatic serial sectioning technique with 15-nm thickness through focused ion beam was employed to observe the three-dimensional structure of exosomes of various sizes. Cryo-transmission electron microscopy revealed the near-to-native structure of exosomes.

Current Status of Automatic Serial Sections for 3D Electron Microscopy

  • Choi, Hyosun;Jung, Min Kyo;Mun, Ji Young
    • Applied Microscopy
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    • 제47권1호
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    • pp.3-7
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    • 2017
  • The automatic equipment for three-dimensional electron microscopy (3DEM) can acquire serial sections of a large sample in a relatively short time, and is especially suitable for the connectomics, which is a field related to understanding the brain structure as a whole. As many results obtained through 3DEM using automatic serial sections have been published in the field of brain research, many researchers continue to apply this technique to various samples. We reviewed the equipment for automatic serial sectioning, the block preparation method, the limitations of 3DEM, and future directions.

Optical Tracking of Three-Dimensional Brownian Motion of Nanoparticles

  • Choi C. K.;Kihm K.D.
    • 한국가시화정보학회지
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    • 제3권1호
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    • pp.3-19
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    • 2005
  • Novel optical techniques are presented for three-dimensional tracking of nanoparticles; Optical Serial Sectioning Microscopy (OSSM) and Ratiometric Total Internal Reflection Fluorescent Microscopy (R-TIRFM). OSSM measures optically diffracted particle images, the so-called Point Spread Function (PSF), and dotermines the defocusing or line-of-sight location of the imaged particle measured from the focal plane. The line-of-sight Brownian motion detection using the OSSM technique is proposed in lieu of the more cumbersome two-dimensional Brownian motion tracking on the imaging plane as a potentially more effective tool to nonintrusively map the temperature fields for nanoparticle suspension fluids. On the other hand, R-TIRFM is presented to experimentally examine the classic theory on the near-wall hindered Brownian diffusive motion. An evanescent wave field from the total internal reflection of a 488-nm bandwidth of an argon-ion laser is used to provide a thin illumination field of an order of a few hundred nanometers from the wall. The experimental results show good agreement with the lateral hindrance theory, but show discrepancies from the normal hindrance theory. It is conjectured that the discrepancies can be attributed to the additional hindering effects, including electrostatic and electro-osmotic interactions between the negatively charged tracer particles and the glass surface.

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Three-dimensional reconstruction of polycrystals using a series of EBSD maps obtained from Dual-beam experiments

  • Kim, MinJi;Son, Youngkyun;Lee, Myeongjin;Jeon, Youngju;Lee, Sukbin
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2016년도 제50회 동계 정기학술대회 초록집
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    • pp.172-172
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    • 2016
  • Dual-beam experiments (Focused ion beam - Orientation mapping microstructure, FIB-OIM) is a widely used experimental tool because this experiments tool available alternates between automated serial sectioning and EBSD with the help of dual beams. We investigated the reconstruction procedure for analysis tool which three-dimensional internal microstructure using Ni superalloy(IN100) and ZrO2. As a results, we observed annealing twin boundary each layer in Ni superalloy(IN100) and fairly isotropic internal microstructure in ZrO2 using marching cubes algorithm. According to these results, this procedure is reconstructed well and we gained ability to arrange the EBSD map and internal microstructure.

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집속이온빔장치와 주사전자현미경을 이용한 박막 트랜지스터 구조불량의 3차원 해석 (Three Dimensional Reconstruction of Structural Defect of Thin Film Transistor Device by using Dual-Beam Focused Ion Beam and Scanning Electron Microscopy)

  • 김지수;이석열;이임수;김재열
    • Applied Microscopy
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    • 제39권4호
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    • pp.349-354
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    • 2009
  • TFT-LCD의 구조불량이 발생한 박막 트랜지스터에 대해서 집속이온빔 가공장치(Dual-beam FIB/SEM)를 이용하여 연속절편법(Serial sectioning)과 일련의 연속적인 2차원 주사전자현미경 이미지를 얻었고, IMOD 소프트웨어를 통해서 3차원 구조구현(3D reconstruction) 연구를 하였다. 3차원 구조구현 결과, Gate막과 Data막이 접합되어 있는 불량이 관찰되었다. 두 막이 접합되어서 ON/OFF 역할을 하는 Gate의 기능이 상실되었고, Data신호는 Drain을 통해서 투명전극에 전류를 공급하여 계속 빛나는 선 불량(line defect)이 발생한 것으로 판단된다. 이 논문의 결과인 집속이온빔 가공장치(Dual-Beam FIB/SEM)를 이용한 3차원 구조구현 연구와 연속절편법, 주사전자현미경 이미지작업, 이미지 프로세싱에 대한 결과는 향후 연구의 기초자료로 활용될 수 있을 것으로 판단된다.

마이크로 X-ray CT를 활용한 알루미늄 개방형 폼의 형상 및 압축 거동 분석 (Analysis of 3D Geometry and Compressive Behavior of Aluminum Open Cell Foam Using X-ray Micro CT)

  • 김영일;김지훈;이종국;김대용
    • 소성∙가공
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    • 제20권7호
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    • pp.518-523
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    • 2011
  • The three dimensional geometries of an aluminum open cell foam before and after uniaxial compressive loading were investigated using the X-ray micro CT(computed tomography). Aluminum 6101-T6 open cell foams of 10, 20, 40 ppi (pore per inch) were considered in this work. After the serial sectioning CT images of aluminum foams were obtained from non-destructive X-ray images, the exact 3D structure were reproduced and visualized with commercial image processing program. The relative density ratio was around the 7.0 to 9.0 range, the unit cells showed anisotropic shapes having the different dimensional ratios of 1.1 to 1.3 between the rise and the transverse directions. The yield stress increased with the relative density ratio and the volumetric strain increased proportionally with compressive strain. The plateau stress in the compressive stress-strain curve was caused by the buckling of ligaments.

New Evaluation System of Cosmetic Effects on Morphology of Skin Surface Using TSRLM with Image Analyser

  • Kim, Jong-Il;Lee, Joa-Hoon;Lee, Yoo-Young;Kim, Chang-Kew
    • 대한화장품학회지
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    • 제16권1호
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    • pp.47-63
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    • 1990
  • Image analyser was used to understand the condition of skin surface and to evaluate the efficacy of cosmetic treatment. It was unsatisfactory to analyse skin surface structure although several methods using image analyser had been presented. We developed the new system composed of image analyser and Tandem Scanning Reflected Light Microscope (TSRLM) having the remarkable optical sectioning property as image input device. By using this new system, we quantitatively measured the change of skin surface, the depth and width of furrow in micron unit, resulted by cosmetic treatments. And also three dimensional image of skin was reconstructed with serial sectioned images, which were captured through TSRLM, for better understanding of the effect of cosmetic treatment. It was found that skin relief was more easily understood and the change of skin surface caused by cosmetic treatment was more accurately measured by using this system. In addition, we was also aware of the possibility of in vivo direct measurement of skin furrow without replica. It was conceivable that our system could be applicable for study of cosmetic effects further.

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하치조신경 절단 후 흰쥐 삼차신경절에서 VIP-IR의 변화 (VIP IMMUNOREACTIVITY IN THE RAT TRIGEMINAL GANGLION AFTER INFERIOR ALVEOLAR NERVE AXOTOMY)

  • 김흥중;박주철;김현섭;문주훈
    • Restorative Dentistry and Endodontics
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    • 제25권2호
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    • pp.225-234
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    • 2000
  • The purpose of this study was to investigate the distribution and fluorescene intensity of vasoactive intestinal polypeptide(VIP) immunoreactive cells in rat trigeminal ganglion after inferior alveolar nerve axotomy. The animals were divided into normal and two experimental groups. The experimental animals were sacrificed at 14th and 28th day after inferior alveolar nerve axotomy. The trigeminal ganglion was removed and immersed in the 4% paraformaldehyde-0.2% picric acid in 0.1M phosphate buffer. Serial frozon sections about $16{\mu}m$ in thickness were cut with a cryostat. The immunofluorescence staining was performed. The rabbit anti-VIP(1 : 8,000) was used as primary antibody and fluorescene isothiocynate(FITC)-conjugated anti-rabbit IgG(1 : 80) as secondary antibody. The slides were observed under confocal laser scanning microscope. Three-dimensional images were constructed from 9 serial images(each $1{\mu}m$ in thickness) made by automatic optical sectioning. Unprocessed optical sections were obtained and stored on a optical disk. Color picture were printed by a video copy processor. The results were as follows; 1. The appearance of VIP immunoreactive cells in the mandibular part of trigeminal ganglion was 8.79${\pm}$1.99% in normal group and 39.16${\pm}$5.62% in 14 days, 16.25${\pm}$2.39% in 28 days after inferior alveolar nerve axotomy groups. 2. The relative fluorescence intensity of VIP immunoreactive cell bodies in the mandibular part of trigeminal ganglion was 134.40${\pm}$10.39 in normal group and 192.88${\pm}$14.06 in 14 days, 143.10${\pm}$5.02 in 28 days after nerve axotomy groups. Therefore, the relative fluorescence intensity of 14 days after nerve axotomy group was 43.3% higher than intensity of normal group. 3. In optical single section analysis of VIP immunoreactive cell bodies, white cell bodies(moderate fluorescence intensity) were the most abundant in normal and 28 days after nerve axotomy groups. Whereas, in 14 days after nerve axotomy group, red cell bodies(high fluorescence intensity) were the most abundant. 4. In optical serial section analysis of VIP immunoreactive cell bodies, red cell bodies(high fluorescence intensity) were observed in a part of the 9 sections of normal and 24 days after nerve axotomy groups. Whereas, red cell bodies were observed in all of the 9 sections of 14 days after nerve axotomy group. 5. The results indicates that number and fluorescence intensity of VIP immunoreactive cells were increased in the mandibular part of trigeminal ganglion following inferior alveolar nerve axotomy.

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실험으로 유도된 구개열 마우스의 안면골 성장에 관한 연구 (A STUDY ON FACIAL BONE GROWTH OF PALATAL CLEFTS EXPERIMENTALLY INDUCED IN MICE)

  • 전윤식
    • 대한치과교정학회지
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    • 제18권2호
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    • pp.329-342
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    • 1988
  • In methods of finding causes for cleft palate, many cases have been studied by investigators using teratogenic agents. Among them, a synthetic agent known as triamcinolone acetonide (TA) was widely used. When this drug was injected into mice during palatogenesis, it induced lowered body weight and a deformed mandible. But many cases have been studied on growth changes, only of the developmental stages of the palate. Therefore, the objective of this study was to evaluate craniofacial growth in experimentally induced cleft palate mice after finishing palatogenesis namely just before birth. Normal, alcohol treated, and TA treated DDY mice were obtained at 18-days of gestation and heads were prepared for serial sectioning in the sagittal plane. The midsagittal sections were photographically enlarged (${\times}40$) and measurements made to asses the amount of growth. The obtained results were as follows. 1. The incidence of cleft palate was 41.2% when TA was injected. 2. The body weight of the cleft palate group was lower than the control group. 3. In the cleft palate group, mandibular length (H-M) was lighter than the control group. 4. In the cleft palate group, degree of staining was not distinct compared to the control group by the double staining method. 5. In the cleft palate group, anteroposterior posture of the tongue tip to facial plane (C-M) was more posterior than the control group. 6. The cause of posterior posture of the tongue tip to facial plane (C-M) in the cleft palate group, was not short and retracted tongue but the mandibular length was increased. 7. The anteroposterior relationship of hyoid cartilage to cranial base was the same in all groups.

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