• Title/Summary/Keyword: confocal laser scanning microscopy (CLSM)

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Visualization of Epidermis and Dermal Cells in ex vivo Human Skin Using the Confocal and Two-photon Microscopy

  • Choi, Sang-Hoon;Kim, Wi-Han;Lee, Yong-Joong;Lee, Ho;Lee, Weon-Ju;Yang, Jung-Dug;Shim, Jong-Won;Kim, Jin-Woong
    • Journal of the Optical Society of Korea
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    • v.15 no.1
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    • pp.61-67
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    • 2011
  • The confocal laser scanning microscopy and two-photon microscopy was implemented based on a single laser source and an objective lens. We imaged and compared the morphology of identical sites of ex vivo human skin using both microscopes. The back-scattering emission from the sample provided the contrast for the confocal microscopy. The intrinsic autofluorescence and the second harmonic generation were used as the luminescence source for the two-photon microscopy. The wavelength of the Ti:Sapphire laser was tuned at 710 nm, which corresponds to the excitation peak of NADH and FAD in skin tissue. The various cell layers in the epidermis and the papillary dermis were clearly distinguished by both imaging modalities. The two-photon microscopy more clearly visualized the intercellular region and the nucleus of the cell compared to the confocal microscopy. The fibrous structures in the dermis were more clearly resolved by the confocal microscopy. Numerous cells in papillary dermal layer, as deep as $100\;{\mu}m$, were observed in both CLSM and two-photon microscopy. While most previous studies focused on fibrous structure imaging (collagen and elastin fiber) in the dermis, we demonstrated that the combined imaging with the CLSM and two-photon microscopy can be applied for the non-invasive study of the population, distribution and metabolism of papillary dermal cells in skin.

Analysis of External Fibrillation of Fiber by Confocal Reflection Microscopy (공초점반사현미경법을 이용한 섬유의 외부소섬유화 분석)

  • Kwon, Ohkyung
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.46 no.2
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    • pp.35-45
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    • 2014
  • Confocal Reflection Microscopy (CRM) was applied to investigate external fibrillation of different types of fibers such as Kajaani reference fiber, Whatman filter fiber, thermomechanical pulp (TMP), and recycled TMP fiber. It was confirmed that the CRM images are created from surface structures of the fiber cell wall. Confocal Laser Scanning Microscopy (CLSM) captured overall shape of the fiber, but minute details of the surface of the fiber were missed. CRM captured the minute details of the fiber surface. From the CRM and CLSM images, it was observed that the CRM images mainly appeared on the fiber surfaces. External fibrillation of the fiber occurs at the fiber surface, not inside the cell wall. Thus, it was concluded that investigation on the external fibrillation of the fiber was possible by utilizing CRM images. A direct qualtitative and quantitative method for analysis of external fibrillation of fiber was demonstrated by utilizing surface area to volume ratio, volume fraction, and roughness calculated from 3-dimensional images reconstructed from stacks of CRM images from the different fibers.

Particle Image Velocimetry of the Blood Flow in a Micro-channel Using the Confocal Laser Scanning Microscope

  • Kim, Wi-Han;Kim, Chan-Il;Lee, Sang-Won;Lim, Soo-Hee;Park, Cheol-Woo;Lee, Ho;Park, Min-Kyu
    • Journal of the Optical Society of Korea
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    • v.14 no.1
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    • pp.42-48
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    • 2010
  • We used video-rate Confocal Laser Scanning Microscopy (CLSM) to observe the motion of blood cells in a micro-channel. Video-rate CLSM allowed us to acquire images at the rate of 30 frames per second. The acquired images were used to perform Particle Image Velocimetry (PIV), thus providing the velocity profile of the blood in a micro-channel. While previous confocal microscopy-assisted PIV required exogenous micro/nano particles as the tracing particles, we employed blood cells as tracing particles for the CLSM in the reflection mode, which uses light back-scattered from the sample. The blood flow at various depths of the micro-channel was observed by adjusting the image plane of the microscope. The velocity profile at different depths of the channel was measured. The confocal micro-PIV technique used in the study was able to measure blood velocity up to a few hundreds ${\mu}m/sec$, equivalent to the blood velocity in the capillaries of a live animal. It is expected that the technique presented can be applied for in vivo blood flow measurement in the capillaries of live animals.

Quantitative Analysis of Pulp fiber Characteristics that Affect Paper Properties (II) (종이의 물성에 영향하는 섬유특성의 정량적 해석(II))

  • 이강진;박종문
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.32 no.2
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    • pp.35-39
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    • 2000
  • Refining is very important process of fibers treatment for proper paper properties. An extent of refining is usually measured by freeness, although freeness gives complicated meanings. One of a direct way of studying the refining effects on pulp fibers is making photomicrographs of beaten fibers. The conventional microscopy like light microscopy(LM) and scanning electron microscopy(SEM) require to preserve the wet structure of pulp fibers morphologically since most of papermaking process is carried out almost entirely in water. Recently developed microscopy, especially confocal laser scanning microscopy(CLSM), offers the possibility of examining fully hydrated pulp fibers. Cross-sectional images of wet pulp fibers are also generated using optical sectioning by CLSM and image analysis in order to verify and quantify the extent of fiber wall swelling indicating the internal fibrillation. At low beating load such as 2.5 kgf, in the same freeness, breaking length is higher than that of high beating load such as 5.6 kgf. fiber wall thickness at low beating load is greater than that at high beating load. This result is accounted for the fact that internal fibrillation in the low beating load was high.

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COMPARATIVE STUDY ON THE EFFICACY OF DIGITAL IMAGING FIBER-OPTIC TRANS-ILLUMINATION AND LASER FLUORESCENCE IN MONITORING THE REMINERALIZATION PROCESS OF INCIPIENT SMOOTH SURFACE ENAMEL LESIONS (Digital Imaging Fiber-Optic Trans-Illumination과 Laser Fluorescence를 이용한 평활면 초기우식증의 재광화에 관한 비교 연구)

  • Hwang, Kyu-Seon;Yoo, Seung-Hoon;Kim, Jong-Soo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.34 no.2
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    • pp.183-191
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    • 2007
  • Through out the world dental caries seems to be decreased as it is difficult to make an accurate diagnosis for dental caries. The traditional diagnostic method which is probing and x-ray taking has many limitations to diagnose the early caries, so there were recommendations for the needs of new equipments such as laser fluorescence(LF), digital imaging fiber-optic trans-illumination(DIFOTI), and quantitative light fluorescence (QLF) which were developed from various study results. Also confocal laser scanning microscopy(CLSM) and ultrasonics are used for research progression. This study is to evaluate whether it is possible to monitor accurately for remineralization amount of enamel surface early caries using DIFOTI or LF After inducing artificial caries to bovine teeth to 10 participants remineralization was enhanced by 0 ppm and 500 ppm fluoride mouth rinse solution for 3 weeks. Then they were cross sectioned and analyzed using gold standard of the lesion depth measured by CLSM. The following results were obtained: 1. The measured percentage of light intensity(luminosity ratio) by DIFOTI increased with remineralization period, and showed significant reverse correlation with lesion depth measured by CLSM (p<0.01). 2. The measurement of laser fluorescence increased with remineralization period, and showed significant correlation with lesion depth measured by CLSM (p<0.01). 3. To the result for CLSM, 500 ppm fluoride mouth rinse group showed rapid rate for decreased tendency of lesion depth than 0 ppm fluoride mouth rinse group. In conclusion DIFOTI system was used to measure accurately for the remineralization amount of early surface caries, it is a very useful equipment to detect precisely the changes for early enamel caries remineralization during treatments.

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Microbial and Physicochemical Monitoring of Granular Sludge During Start-up of Thermophilic UASB Reactor

  • Ahn, Yeong-Hee;Park, Sung-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.378-384
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    • 2003
  • Mesophilically-grown granular sludge seeded in thermophilic UASB reactor was monitored to better understand the start-up process of the reactor. The reactor was fed with a synthetic wastewater containing glucose. As COD loading rate increased stepwise, methane production rate increased. Maximum values of COD removal efficiency (95%) and methane production rate (5.3 l/day) were achieved by approximately day-80 and remained constant afterward. However, physicochemical and microbial properties of granules kept changing even after day-80. Specific methanogenic activity (SMA) was initially negligible, and increased continuously until day-153 and remained constant afterward, showing the maximum value of $1.51{\pm}0.13\;g\;CH_4-COD/g$ VSS/day. Deteriorated settling ability of granules recovered the initial value by day-98 and was maintained afterward, as determined by sludge volume index. Initially reduced granule size increased until day-126, reaching a plateau of 1.1 mm. Combined use of fluorescence in situ hybridization and confocal laser scanning microscopy (CLSM) allowed to localize families of Methanosaetaceae and Merhanosarcinaceae in granules with time Quantitative analyses of CLSM images of granule sections showed abundance patterns of the methanogens and numerical dominance of Methanosaeta spp. throughout the start-up period. The trend of SMA agreed well with abundance patterns of the methanogens.

Hanji Manufacturing from Bast Fibers of Kenaf, Hibiscus cannabinus (양마의 인피섬유를 이용한 한지제조)

  • Cho, Nam-Seok
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.40 no.4
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    • pp.1-9
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    • 2008
  • The utilization of non-woody fibers with the fast growing annual plants has occurred in the paper industry to replace wood and preserve environment of the earth. The non-woody fibers generally used for papermaking are paper mulberry, gampi, manila hemp, rice straw, bamboo, and coton linter etc.. Recently Kenaf has been spot-lighted for the same application. Kenaf is an annual plant of Hibiscus species of Malvaceae family. Kenaf, a rapid growing and high harvesting non-woody fiber plant, was identified as one of the promising fiber sources for the production of paper pulp. This study was carried out to investigate the pulping characteristics of Kenaf bast fiber for Hanji (traditional Korean paper) manufacturing by different pulping methods, such as alkali, alkali-peroxide and sulfomethylated pulpings. It was possible to make superior grade of Hanji. Especially sulfomethylated pulping was resulted in superior pulp in terms of higher yields and qualities in comparison to those of the other pulping methods. Hanji from sulfomethylated pulp was shown the highest brightness of over 60% and higher sheet strength. In addition, the morphological features of pulp fibers (pulp compositions) affect to the sheet properties. Therefore the effect of fiber distribution index(FDI) which was calculated from the data of Confocal laser scanning microscopy(CLSM) on the sheet properties of Kenaf Hanji was also discussed.

Droplet Geometry and Its Volume Analysis (기름방울 형상 및 그 체적 분석법)

  • Yoon, Moon-Chul
    • Tribology and Lubricants
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    • v.24 no.6
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    • pp.320-325
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    • 2008
  • The recent industrial application requires technical methods to get the cutting fluid droplet surfaces in particular from the viewpoint of topography and micro texture. To characterize the surface topography of droplet, the combination of the confocal laser scanning microscope (CLSM) and wavelet filtering is well suited for obtaining the droplet geometry encountered in tribological research. This technique indicates a better agreement in obtaining an appropriate droplet surface obtained by the CLSM over a detail range of surface accuracy (resolution: $2{\mu}m$). And the results allow an excellent accuracy in a measurement of a droplet surface. The combination of extended focal depth measurement configured and multi-scale wavelet filtering has proven that it can construct a droplet surface in a successive and accurate way. A multi-scale approach of wavelet filtering was developed based on the decomposition and reconstruction of droplet surface by 2D wavelet transform using db9 (a mother wavelet of daubechies). Also this technique can be extended to characterize the quantification of droplet properties and other field in a wide range of scales. Finally this method is verified to be a better droplet surface modeling in a micro scale arising in a mist machining.

Clinical Microscopy: Performance, Maintenance and Laser Safety (임상에서의 현미경: 작동, 유지보수 및 레이저 안전)

  • Lee, Tae Bok
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.2
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    • pp.125-133
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    • 2019
  • A microscope is the fundamental research and diagnostic apparatus for clinical investigation of signaling transduction, morphological changes and physiological tracking of cells and intact tissues from patients in the biomedical laboratory science. Proper use, care and maintenance of microscope with comprehensive understanding in mechanism are fully requested for reliable image data and accurate interpretation for diagnosis in the clinical laboratory. The standard operating procedure (SOP) for light microscopes includes performance procedure, brief information of all mechanical parts of microscopes with systematic troubleshooting mechanism depending on the laboratory capacity. Maintenance program encompasses cleaning objective, ocular lenses and inner optics; replacement and calibration of light source; XY sample stage management; point spread function (PSF) measurement for confocal laser scanning microscope (CLSM); quality control (QC) program in fluorescent microscopy; and systematic troubleshooting. Laser safety is one of the concern for medical technologists engaged in CLSM laboratory. Laser safety guideline based on the laser classification and risk level, and advisory lab wear for CLSM users are also expatiated in this overview. Since acquired image data presents a wide range of information at the moment of acquisition, well-maintained microscopes with proper microscopic maintenance program are impulsive for its interpretation and diagnosis in the clinical laboratory.

CLSM [Confocal Laser Scanning Microscope] Observation of the Surface Roughness of Pressurized Rock Samples During Freeze/Thaw Cycling

  • Kim, Hye-jin;Choi, Junghae;Chae, Byung-gon;Kim, Gyo-won
    • The Journal of Engineering Geology
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    • v.25 no.2
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    • pp.165-178
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    • 2015
  • Physical and chemical weathering degrades rock, affecting its structural properties and thus the stability of stone buildings or other structures. Confocal laser scan microscopy (CLSM) is used here to observe temporal changes in the surface roughness of rock samples under simulated accelerated weathering. Samples were pressurized to 50, 55, or 70 MPa using a pressure frame, and subjected to freeze/thaw cycling controlled by a thermostat. The temperature was cycled from -20℃ to 40℃ and back. After each 20 cycles, CLSM was used to assess the change in surface roughness, and roughness factors were calculated to quantify the progression of the surface condition over time. Variations in cross-section line-roughness parameters and surface-roughness parameters were analyzed for specific parts of the sample surfaces at 5× and 50× magnification. The result reveals that the highest and lowest values of the roughness factors are changed according to elapsed time. Freezing/thawing at high pressure caused larger changes in the roughness factor than at low pressure.