• 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.

• Journal of the korean academy of Pediatric Dentistry
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• v.28 no.1
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• pp.25-31
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• 2001

The purpose of this study was to investigate the distribution and fluorescence intensity of vasoactive intestinal polypeptide immunoreactive (VIP-IR) cells in rat trigeminal ganglion following pulp extirpation of rat mandibular molar. The animals were divided into control group(n=6) and experimental group(n=6). The experimental animals were sacrificed at 14 days after pulp extirpation. The trigeminal ganglion was removed and immersed in the 4% paraformaldehyde in 0.1M phosphate buffer. Serial frozen sections about $20{\mu}m$ 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 (CLSM). Unprocessed optical sections were obtained and stored on a optical disk. Color pictures were printed by a video copy processor. The results were as follows; 1. The positive ratio of VIP-IR cells in mandibular part of trigeminal ganglion were 7.40% in control group and 28.42% in experimental group(14 days after pulp extirpation). 2. The relative fluorescence intensity of VIP-IR cells in mandibular part of trigeminal ganglion were 87.78 in control group and 138.65 in experimental group. The relative fluorescence intensity of experimental group was 58% higher than that of control group. 3. In optical serial section analysis of VIP-IR cells of experimental group, most of the 9 sections showed high fluorescence intensity. At high magnification, axons of the experimental group displayed greater VIP-IR than in the control group, and the positive cells were mainly of medium size. The result indicate that number and fluorescence intensity of VIP-IR cells were increased in the mandibular part of trigeminal ganglion following pulp extirpation of mandibular molar, and it suggests that VIP could play a role in processing of nociception.

• Neonatal Medicine
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• v.15 no.1
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• pp.22-31
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• 2008

Purpose : Bronchopulmonary dysplasia (BPD) is characterized by arrested vascular and alveolar growth in the premature lung. Considering the consequences of arrested lung growth, the idea of administering bone marrow cells to enhance the inborn repair mechanism is promising as this may reduce the morbidity and mortality of BPD. We followed enhanced green fluorescent protein (EGFP)-labeled bone marrow cells (BMC) injected intraperitoneally into non-EGFP mice in order to determine their fate after transplantation. Methods : An angiogenesis inhibitor, SU1498, was injected subcutaneously on day 3 in non-EGFP C57BL/6 newborn mice to create a model of arrested alveolar development. On the following day, $1{\times}10^6$ BMCs isolated from major histocompatibility complex (MHC)- matched syngenic EGFP mice were injected intraperitoneally to non-EGFP BPD mice. Morphometric analysis, immunostaining, and confocal microscopy were performed to determine the fate of EGFP-positive stem cells in the injured lung. Results : SU1498 injection reduced alveolar surface area and mean alveolar volume in newborn mice. BMC injection resulted in recovery of lung structure comparable to controls. EGFP-positive BMCs were identified in the lungs of the recipient mice after intraperitoneal injection. The injected EGFP cells were co-stained with endothelial and epithelial cells of the developing lung as determined by confocal microscopy. Conclusion : Our results illustrated that EGFP-positive BMCs engrafted and trans-differentiated into epithelial and endothelial cells after intraperitoneal injection in a mouse model of arrested alveolar development.

• Restorative Dentistry and Endodontics
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• v.33 no.1
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• pp.1-8
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• 2008

The purpose of this study was to evaluate the effect of a new resin monomer, filler size and polishing technique on the surface roughness of composite resin restorations using confocal laser scanning microscopy. By adding new methoxylated Bis-GMA (Bis-M-GMA, 2,2-bis[4-(2-methoxy-3-methacryloyloxy propoxy) phenyl] propane) having low viscosity, the content of TEGDMA might be decreased. Three experimental composite resins were made: EX1 (Bis-M-GMA/TEGDMA = 95/5 wt%, 40 nm nanofillers); EX2 (Bis-M-GMA/TEGDMA = 95/5 wt%, 20 nm nanofillers); EX3 (Bis-GMA/TEGDMA = 70/30 wt%, 40 nm nanofillers). Filtek Z250 was used as a reference. Nine specimens (6 mm in diameter and 2 mm in thickness) for each experimental composite resin and Filtek Z250 were fabricated in a teflon mold and assigned to three groups. In Mylar strip group, specimens were left undisturbed. In Sof-lex group, specimens were ground with #1000 SiC paper and polished with Sof-lex discs. In DiaPolisher group, specimens were ground with #1000 SiC paper and polished with DiaPolisher polishing points. The Ra (Average roughness), Rq (Root mean square roughness), Rv (Valley roughness), Rp (Peak roughness), Rc (2D roughness) and Sc (3D roughness) values were determined using confocal laser scanning microscopy. The data were statistically analyzed by Two-way ANOVA and Tukey multiple comparisons test (p = 0.05). The type of composite resin and polishing technique significantly affected the surface roughness of the composite resin restorations (p < 0.001). EX3 showed the smoothest surface compared to the other composite resins (p < 0.05). Mylar strip resulted in smoother surface than other polishing techniques (p < 0.05). Bis-M-GMA. a new resin monomer having low viscosity, might reduce the amount of diluent, but showed adverse effect on the surface roughness of composite resin restorations.

• Proceedings of the Korean Society For Composite Materials Conference
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• 2001.05a
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• pp.201-204
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• 2001

A Confocal Laser Scanning Microscope (CLSM) is applied to determine three-dimensional fiber orientation states in injection-molded short fiber composites. Since the CLSM optically sections the composites, more than two planes either on or below the surface of composites can be obtained. Therefore, three dimensional fiber orientation states are determined without destruction. To predict the orientation states, velocity and temperature fields are calculated by using a hybrid FEM/FDM method. The change of orientation state during packing stage is also considered by employing a compressible Hele-Shaw model. The predicted orientation states show good agreement with measured ones. However, some differences are found at the end of cavity. They may result from other effects, which are not considered in the numerical analysis.

• Journal of Pharmacopuncture
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• v.11 no.2
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• pp.5-12
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• 2008

Objective In this article, we report on the intradermal Alcian blue staining method for tracing the meridians of acupuncture. Methods 1% Alcian blue solution was injected into acupoints by using a 0.5mL insulin syringe with a 31-gauge needle, then the skin was incised and was observed under a stereoscopic microscope. The specimens were examined by using immunohistochemical methods and were observed under a confocal laser scanning microscope. Results A threadlike structure, which was visualized with Alcian blue, existed in dermis layer and proceeded to hypodermis. In this structure, characteristic alignments of rod- shaped nuclei and $1-2{\mu}m$ sized DNA granules were observed. Furthermore, abundant blood capillary plexuses, peripheral nerve endings, and a corpuscle-like structure(about $300{\mu}m$ in diameter) were visualized in the skin tissues of acupoints. Conclusion It was concluded that the specific threadlike and corpuscle-like structures corresponded to superficial Bonghan duct and corpuscle, respectively.

• Journal of the Korean Society of Manufacturing Technology Engineers
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• v.18 no.3
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• pp.248-254
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• 2009

The worn crater wear geometry of coated tools after machining has been configured by using Confocal Laser Scanning Microscopy(CLSM) and the Wavelet-based filtering technique. The CLSM can be well suited to construct the three-dimensional crater wear on the rake surfaces of coated tips. However, The raw heightness data of HEI(height encoded image) acquired by CLSM must be filtered due to the electronic and imaging noise occurring in constructing the crater image. So the Wavelet-based filtering algorithm is necessary to denoise the shape features in a micro scales so as to realize accurate crater wear topography analysis. The crater wear patterns filtered enable us to predict the crater wear shape in order to study the tool wear evolution. The study shows that the technique by combining the CLSM and Wavelet-based filtering is an excellent one to obtain the geometries of worn tool rake surfaces over a wide range of surface resolution in a micro scale.

• Transactions of the Korean Society of Mechanical Engineers B
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• v.30 no.9 s.252
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• pp.834-841
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• 2006

In the present wort the Laser-induced Fluorescence (LIF) technique and Confocal Laser Scanning Microscopy (CLSM) have been combined to measure the temperature distribution across a micro-scale liquid layer as a direct and non-invasive method. Only the fluorescent light emitted from a very thin volume around a focal plane can be selectively detected, and it enables us to measure the liquid temperatures even at the close vicinity of the walls. As an experimental verification, a test section consists of two flat plates (for heating and cooling, respectively) separated by about 240 microns was made, and the methanol mixed with a temperature-sensitive dye, Rhodamine B, was filled in the gap between them. The measured temperature distribution across the gap showed good linearity, which is a typical characteristic of conduction heat transfer through a thin liquid layer. In result, the CLSM-LIF technique proposed in the present study was found to be a promising method to measure the local temperatures in the liquid flow field in microfluidic devices.

• Journal of the Semiconductor & Display Technology
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• v.7 no.1
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• pp.11-16
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• 2008

Confocal scanning microscopy is a widely used technique for three dimensional measurements because it is characterized by high resolution, high SNR and depth discrimination. Generally an image is generated by moving one optical probe that satisfies the confocal condition on the specimen. Measurement speed is limited by movement speed of the optical probe; scanning speed. To improve measurement speed we increase the number of optical probes. Specimen region to scan is divided by optical probes. Multi-point information each optical probe points to can be obtained simultaneously. Therefore image acquisition speed is increased in proportion to the number of optical probes. And multiple optical probes from red, green and blue laser sources can be used for color imaging and image quality, i.e., contrast, is improved by adding color information by this way. To conclude, this technique contributes to the improvement of measurement speed and image quality.

• Restorative Dentistry and Endodontics
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• v.26 no.1
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• pp.1-8
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• 2001

Dental Caries which has high prevalence rate, accounts for majority of dental diseases. Many treatment and preventive treatment has been developed, thereby reducing the prevalence rate, but in our country, fluoridization has not spread widely yet, so prevention has not been done satisfactorily. When dental caries progresses, irreversible damage of tooth structure occurs. In initial dental caries, demineralizing tooth structure can be remineralized, so restorative treatment is unnecessary. In this study, 20 teeth restored with composite resin without fluoride release were used and divided into two groups. Incipient dental caries were artificially made and demineralization procedure was done for 1 and 2 weeks, for each group. Changes in mineral contents around the margins were analysed with confocal laser scanning microscope. The results were as follow. 1. Both total fluorescence of the lesion and average fluorescence of the lesion of remineralized samples decreased compared to demineralizing state. (p＜0.01) 2. Confocal laser scanning microscopy can be used in quantitative analysis of mineral change. In result, confocal laser scanning microscopy can be used in quantitative analysis of mineral change and it could be used in many different fields of dentistry in the future.