• Title/Summary/Keyword: Confocal Fluorescence Image

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Spectroscopic Comparison of Photo-oxidation of Outside and Inside of Hair by UVB Irradiation (자외선B 조사에 의한 모발 외부와 내부의 광산화에 관한 분광학적 비교)

  • Ha, Byung-Jo
    • Applied Chemistry for Engineering
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    • v.31 no.2
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    • pp.220-225
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    • 2020
  • Hair is made of proteins containing various amino acids. Ultraviolet (UV) radiation is believed to be responsible for the most damaging effects of sunlight, and also plays an important role in hair aging. The purpose of this study was to investigate the changes in morphological and chemical structures after ultraviolet B (UVB) irradiation of human hair. The UVB-irradiated hair showed characteristic morphological and structural changes, compared to those of the normal hair. The result from a scanning electron microscope (SEM) equipped with an energy dispersive X-ray diffractometer (EDX) showed that the scale of UV-irradiated hair appeared to be rough and the amount of oxygen element was higher than that of the normal hair. Fluorescence and three dimensional (3D) topographical images were obtained by a confocal laser scanning microscope (CLSM). In 3D images, the green emission intensity of normal hair was much higher than that of fluorescing UVB-irradiated hair. The intensity of green emission reflects the intrinsic fluorescence of hair protein. Also, a fluorescent imaging method using fluorescamine reagent was used to identify the free amino groups resulting from a peptide bond breakage in UVB-irradiated hair. Strong blue fluorescence of UVB-irradiated hair, which indicates a very high level of amino groups, was observed by CLSM. Therefore, the fluorescamine as an extrinsic fluorescence could provide a useful tool to identify the peptide bond breakage in UVB-irradiated hair. Infrared image mapping was also employed to assess the cross-sections of normal and UVB-irradiated specimens to examine the oxidation of disulfide bonds. The degree of peak areas with strong absorbance for the disulfide mono-oxide was spread from the outside to the inside of hair. The spectroscopic techniques used alone, or in combination, launch new possibilities in the field of hair cosmetics.

Fluorescent Method for Observing Intravascular Bonghan Duct (형광염색을 이용한 혈관내봉한관의 관찰)

  • Lee, Byung-Cheon;Baik, Ku-Youn;Johng, Hyeon-Min;Sung, Baeck-Kyoung;Soh, Kyung-Soon;Kang, Dae-In;Soh, Kwang-Sup
    • Journal of Pharmacopuncture
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    • v.8 no.3
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    • pp.5-9
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    • 2005
  • Observation of intra-vascular threadlike structures in the blood vessels of rats is reported with the images by differential interference contrast microscope, and fluorescence inverted microscope of the acridine-orange stained samples. The confocal microscope image and the hematoxylin-eosin staining revealed the distinctive pattern of nuclei distribution that clearly discerned the threadlike structure from fibrin, capillary, small venule, arteriole, or lymph vessel. Physiological function of the intra-vascular thread in connection with acupuncture is discussed. Especially, this threadlike duct can be a circulation path for herb-liquid flow, which may provide the scientific mechanism for therapeutic effect of herbal acupuncture.

Diagnostic Utilization of Laser Fluorescence for Resin Infiltration in Primary Teeth (유치의 레진침투법을 위한 레이저 형광법의 진단적 활용)

  • Park, Soyoung;Jeong, Taesung;Kim, Jiyeon;Kim, Shin
    • 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.

Characterization of the Expression of PKCα(Isoform) in DMH-induced Vascular Endothelial Proliferation (DMH에 의한 비정상적인 혈관 내피세포의 증식에서 Protein Kinase C 동종효소 Alpha 단백발현의 특성)

  • Nam, Su Bong;Bae, Yong Chan;Park, Suk Young;Choi, Soo Jong
    • Archives of Plastic Surgery
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    • v.34 no.6
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    • pp.679-684
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    • 2007
  • Purpose: DMH(1,2-dimethylhydrazine) has been known to induce vascular neoplasm such as malignant endothelioma in animal experiment, through induction of abnormal proliferation of HUVECs. In our previous studies, 11 types of PKC isoenzymes were determined by RT-PCR and the expression of $PKC{\alpha}$, and ${\mu}$ was more prominent than other PKC isoenzymes in the DMH-treated group. However, this result was not based on objective assessment. In this study, we further evaluated the role of $PKC{\alpha}$ on the DMH-induced abnormal proliferation of HUVECs by two different methods to identify its presence with high relevance in objective view. $PKC{\mu}$ will be investigated in further study. Methods: The study was conducted with the cultured HUVECs group(control) and the $0.75{\times}10^{-9}M$ DMH-treated group. After processing protein extraction in 0 and 24 hour, extracted protein was treated of quantitative test through BCA protein assay. In the western blot analysis, electrophoresis was performed in the order of gel preparation, sample preparation, and gel running. Electrotransfer to nitrocellulose membrane and reaction with antibody were done. Detection of $PKC{\alpha}$ was achieved through "Gel Image Analysis System". In the fluorescence immunocytochemical analysis, the grading of radiance of the intracellular $PKC{\alpha}$ particles was detected with confocal microscope after treating with primary and fluorescent secondary antibody in 0 and 24 hours. Results: The Western blot analysis showed increased $PKC{\alpha}$ expression from the specimen obtained in 24 hour of the DMH treatment group when compared to those in control group. Under confocal fluorescence microscope, the emitting radiance in the DMH treated group was brighter at 24 hours as well. Conclusion: We believe that $PKC{\alpha}$ plays a role in DMH-induced abnormal proliferation of the vascular endothelium, which may provide insights in understanding the vascular neoplasm.

3D Shape Reconstruction using the Focus Estimator Value from Multi-Focus Cell Images (다초점 세포 영상으로부터 추정된 초점 값을 이용한 3차원 형태 복원)

  • Choi, Yea-Jun;Lee, Dong-Woo;Kim, Myoung-Hee;Choi, Soo-Mi
    • 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.

Aluminum toxicity-induced alterations of root proteome in wheat seedlings

  • Oh, Myeong Won;Roy, Swapan Kumar;Cho, Kun;Cho, Seong-Woo;Park, Chul-Soo;Chung, Keun-Yook;Choi, Jong-Soon;Woo, Sun-Hee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.127-127
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    • 2017
  • Aluminum is the most abundant metallic element in the Earth's crust and considered as the most limiting factor for plant productivity in acidic soils. The inhibition of root growth is recognized as the primary effect of Al toxicity. Seeds of wheat cv. Keumkang (Korean cultivar) were germinated on petridish for 5 days and then transferred hydroponic apparatus which was treated with $0{\mu}M$ $AlCl_3$ (control), $100{\mu}M$ $AlCl_3$ and $150{\mu}M$ $AlCl_3$ for 5 days. The length of roots, shoots and fresh weight of wheat seedlings were decreased under aluminum stress. The concentrations of $K^+$, $Mg^{2+}$ and $Ac^{2+}$ were decreased whereas $Al^{3+}$ and $P_2O_5{^-}$ concentration was increased under aluminum stress. Using confocal microscopy, the fluorescence intensity of aluminum was increased with morin staining. In this study, a proteome analysis was performed to identify proteins, which is responsible to aluminum stress in wheat roots. In 10-day-old seedlings, proteins were extracted from roots and separated by 2-DE, stained by CBB. Using image analysis, a total of 47 differentially expressed protein spots were selected, whereas 19 protein spots were significantly up-regulated such as s-adenosylmethionine, oxalate oxidase, malate dehydrogenase, cysteine synthase, ascorbate peroxidase and 28 protein spots were significantly down-regulated such as heat shock protein 70, o-methytransferase 4, enolase, amylogenin by aluminum stress following protein spots analyzed by LTQ-FTICR mass spectrometry. The results provide the global picture of Al toxicity-induced alterations of protein profiles in wheat roots, and identify the Al toxicity-responsive proteins related to various biological processes that may provide some novel clues about plant Al tolerance.

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