BACKGROUND: This study aimed to evaluate the adhesion of Acanthamoeba trophozoites on cosmetic contact lenses (CLs) with and without CL care multipurpose solution (MPS) treatment. METHODS: Acanthamoeba lugdunensis L3a trophozoites were inoculated onto disks trimmed from CLs: 1-day Acuvue moist, 1-day Acuvue define, Acuvue 2, and Acuvue 2 define. After 18-hour inoculation, the number of adherent trophozoites was counted under phase contrast microscopy. The effects of MPS, Opti-Free Express, soaking CLs for 6 hours, on Acanthamoeba adhesion were analyzed. Scanning electron microscopic examination was performed for assessment of Acanthamoeba attached on the lens surface. RESULTS: Acanthamoeba trophozoites showed greater adhesion to cosmetic CL (P = 0.017 for 1-day CL and P = 0.009 for 2-week CL) although there was no significant difference between the types of cosmetic CL. On all lenses, the number of adherent Acanthamoeba was significantly reduced after treatment with MPS (P < 0.001 for 1-day Acuvue moist, P = 0.046 for 1-day Acuvue define, P < 0.001 for Acuvue 2, and P = 0.015 for Acuvue 2 define), but there was still significant difference between conventional and cosmetic CLs (P = 0.003 for 1-day CL and P < 0.001 for 2-week CL, respectively). More attachment of Acanthamoeba was observed on colored area and the acanthopodia of Acanthamoeba was placed on the rough surface of colored area. CONCLUSION: Acanthamoeba showed a greater affinity for cosmetic CL and mostly attached on colored area. Although MPS that contained myristamidopropyl dimethylamine reduced the adhesion rate, there was a significant difference between conventional and cosmetic CLs.
Acanthamoeba ; Contact Lenses ; Microscopy, Phase-Contrast ; Trophozoites

Dendritic cells (DCs) are professional antigen presenting cells of the immune system. They can be generated in vitro from peripheral blood monocytes supplemented with GM-CSF, IL-4 and TNF alpha. During induction, DCs will increase in size and acquire multiple cytoplasmic projections when compared to their precursor cells such as monocytes or haematopoietic stem cells which are usually round or spherical. Morphology of DCs can be visualized by conventional light microscopy after staining or phase-contrast inverted microscopy or confocal laser scanning microscopy. In this report, we described the morphological appearances of DCs captured using the above-mentioned techniques. We found that confocal laser scanning microscopy yielded DCs images with greater details but the operating cost for such a technique is high. On the other hand, the images obtained through light microscopy after appropriate staining or phase contrast microscopy were acceptable for identification purpose. Besides, these equipments are readily available in most laboratories and the cost of operation is affordable. Nevertheless, morphological identification is just one of the methods to characterise DCs. Other methods such as phenotypic expression markers and mixed leukocyte reactions are additional tools used in the characterisation of DCs.
Dendritic Cells/*cytology ; Microscopy, Confocal ; Microscopy, Phase-Contrast

This commentary presents the regulatory backgrounds and development of the national proficiency testing (PT) scheme on asbestos analysis in the Republic of Korea. Since 2009, under the amended Occupational Safety and Health Act, the survey of asbestos in buildings and clearance test of asbestos removal works have been mandated to be carried out by the laboratories designated by the Ministry of Employment and Labor (MOEL) in the Republic of Korea. To assess the performance of asbestos laboratories, a PT scheme on asbestos analysis was launched by the Korea Occupational Safety and Health Agency (KOSHA) on behalf of the MOEL in 2007. Participating laboratories are evaluated once a year for fiber counting and bulk asbestos analysis by phase contrast microscopy and polarized light microscopy, respectively. Currently, the number of laboratory enrollments is > 200, and the percentage of passed laboratories is > 90. The current status and several significant changes in operation, sample preparations, and statistics of assigning the reference values of the KOSHA PT scheme on asbestos analysis are presented. Critical retrospect based on the experiences of operating the KOSHA PT scheme suggests considerations for developing a new national PT scheme for asbestos analysis.
Asbestos* ; Employment ; Korea* ; Microscopy, Phase-Contrast ; Microscopy, Polarization ; Occupational Health ; Reference Values ; Republic of Korea

The in vitro growth pattern of the human retinal pigment epithelial cell was carried out from enucleated eye. The cultured cell was observed serially under phase contrast microscopy and stained for light microscopy. The cultured retinal pigment epithelium no longer resembled the original pattern of the retinal pigment epithelium and lost pigment granules as tissue culture progressed. This work has shown that the retinal pigment epithelium was viable and possessed proliferative potency for up to 36 days after seeding without subculture.
Cells, Cultured ; Epithelial Cells ; Humans* ; Microscopy ; Microscopy, Phase-Contrast ; Retinal Pigment Epithelium* ; Retinaldehyde*

PURPOSE: The purpose of this study is to evaluate the feasibility of phase contrast X-ray microtomography and microradiography, using a polychromatic synchrotron X-ray, for analysis of the mouse lung microstructure. MATERIALS AND METHODS: Normal mice were used for experiments. Some of the mouse lungs were prepared by the lung fixation-inflation method. The resulting sponge-like inflated lung samples were used for microtomography. The remaining mouse lungs were cut into 10 um sections and were used for microradiography and optical microscopic correlation. The experiments on mouse lung samples were performed at the 7B2 beamline of the Pohang Light Source in Korea. RESULTS: Phase contrast X-ray microtomography of inflated lung samples showed individual alveolar structure on 3-D reconstruction. Phase contrast microradiographs of thin lung samples showed microstructure of lung, such as alveoli and bronchioles, and were well correlated with optical microscopic images. CONCLUSIONS: The results indicate that the phase contrast X-ray microtomography and microradiography using polychromatic synchrotron X-ray is feasible for evaluation of microstructure of the lung.
Animals ; Lung/*cytology/*radiography ; Mice ; Microscopy/*methods ; Microscopy, Phase-Contrast ; X-Ray Microtomography/*methods

BACKGROUND: Recently, G1 cells, characterized by distinctive doughnut-like shape with blebs have been reported as a reliable marker for glomerular hematuria. We investigated the validity of the urinary G1 cells in distingushing glomerular from non-glomerular hematuria. In addition, we evaluate the influence of urine osmolality, pH and proteinuria on dysmorphic erythrocytes and G1 cells. METHODS: One hundred and twenty patients with hematuria including 60 glomerular (GH) and 60 non- glomerular hematuria (NGH) were examined. The percentage of urinary dysmorphic erythrocytes and G1 cells using phase-contrast microscopy was determined. Urine osmolality, pH, and spot urine protein/ creatinine ratio were examined. RESULTS: The proportion of G1 cells differed significantly between the two group (7.8+/-16.0% in GH vs. 0% in NGH, p<0.05). At the cut-off value of 50 % dysmorphic erythrocytes, the sensitivity and specificity for the detection of GH was 88.3% and 93.3%, respectively. At the cut-off value of 1% G1 cells, sensitivity and specificity were 60.0% and 100%, respectively. When both of 50% dysmorphic erythrocytes and 1% G1 cells were considered as the cut-off value, the sensitivity and specificity were 91.0% and 100%, respectively. There was a significant difference in the percentage of dysmorphic erythrocytes and G1 cells at different urine pH. There was a significant correlation between urine osmolality and dysmorphic erythrocytes (r=0.41, p< 0.05), but not for G1 cells. No significant correlations were observed between G1 cells and proteinuria or pH. CONCLUSION: Evaluation of both urinary G1 cell and dysmorphic erythrocytes at the same time could improve the diagnostic value for differentiating glomerular hematuria.
Blister ; Creatinine ; Erythrocytes ; Hematuria* ; Humans ; Hydrogen-Ion Concentration ; Microscopy, Phase-Contrast ; Osmolar Concentration ; Proteinuria ; Sensitivity and Specificity

PURPOSE: To investigate the differentiation of lens epithelial cells (LECs) to lens fiber, and the transdifferentiation of LECs to fibroblast in capsular bag culture. METHODS: After observing the changes of LECs by using phase-contrast microscopy, we observed a cross section of capsular bag by using light microscope (LM) and electron microscope (EM). In addition, the expressions of alpha A-crystallin, a marker of differentiation of LEC to lens fiber, and of alpha-smooth muscle actin, a marker of LEC to fibroblast, were examined during the culture period by western blot. RESULTS: On phase-contrast microscopy, 7 to 14 days after culture, the portion of LECs was gradually elongated and cytoplasm became transparent, so that the differentiation resembled lens fiber. One to 7 days after culture, the portion of LECs changed to spindle shape and the transdifferentiation resembled fibroblast. LM and EM observations indicated that changes of each LEC were lens fiber, and fibroblast. According to Western blot, the expression of alpha A-crystallin was increased by 10 days after culture. The alpha-smooth muscle actin showed an increased expression 10 to 30 days after culture. CONCLUSIONS: From the capsular bag model, we observed the resemblances of the differentiation and transdifferentiation of LECs with lens fiber and fibroblast.
Actins ; alpha-Crystallin A Chain ; Blotting, Western ; Cytoplasm ; Epithelial Cells* ; Fibroblasts ; Microscopy, Phase-Contrast

BACKGROUND: In the skin, the major stimulus for cutaneous pigmentation is ultraviolet radiation. The most important physiologic role of melanin is protection against harmful UV radiation to skin. It is known there are some differences in melanization between a single and multiple exposures of UVB, in vivo. Little if known about the functions of the melanocyte alone in cutaneous pigmentation after ultraviolet exposure, because of the complexity of interactions in the whole epidermis. OBJECTIVE: To investigate the effects of multiple exposures at various dosages of UVB, and to compare the effect of UVB in multiple divided exposures with a single exposure at the same total dosage of UVB on proliferation and melanization in cultured human melanocyte. METHODS: Melanocytes were cultured by modified TIC medium. The melanoctes were exposed daily for three consecutive days to UVB at 2, 4, 8 and 16 mJ/cm2and a single exposure at 24 mJ/cm2. The morphologic changes were examined by phase contrast microscopy. The melanocytes were counted by hemocytometer and melanin contents were assayed by spectrophotometer. RESULTS: 1. The effects of multiple UVB exposures: 1) The morphologic changes were as follows: With three time exposures at a dosage of 8 mJ/cm2, themelanocytes enlarged in size, and elongated their dendrites slightly; with three time exposures at a dosage of 16 mJ/cm2, enlargement in sized and elongation of dendrited were more significant. 2) With three time exposures at dosages of 2 nd 4 mJ/cm2, the proliferation of melanocytes was stiumlated significantly(p<0.05). However, with three time exposures at dosages of 8 and 16 mJ/cm2the proliferation was inhibited(p<0.05). 3) With three time exposures at dosages of 2 and 4 mJ/cm2, the melanin contents were decreased. However, with three tiem exposures at a dosage of 16 mJ/cm2, the melanin contents were highly increased(p<0.01). 2. The comparison between multiple divided exposures and a single exposure at the same toal dosage of UVB: 1) There were no morphologic differences of dendrities between with three time exposures at a dosage of 8 mJ/cm2 and with a single exposure at a dosage of 24 mJ/cm2. However enlarged melanocytes were more numerous with a single exposure. 2) The proliferation of melanocytes was more inhibited with a single exposure than with multiple divided exposures(p<0.05). 3) The melanin contents were more increased with a single exposure than with multiple divided exposures(p<0.05). CONCLUSION: With multiple exposures at lower dosages of UVB, the proliferation of melanocytes was stimulated, and melanization was decreased. However, with multiple exposures at higher dosages of UVB, the proliferation was inhibited, and melanization was increased. At the same total dosage of UVB, the proliferation was more inhibited, and the melanization was more increased with a single exposure than with multiple divided exposures.
Dendrites ; Dermatitis, Irritant ; Epidermis ; Humans* ; Melanins ; Melanocytes* ; Microscopy, Phase-Contrast ; Pigmentation ; Skin ; Tics

PURPOSE: To investigate the effects of two antimetabolites, mitomycin C (MMC) and 5-fluorouracil (5-FU), on proliferation of cultured human nasal mucosa fibroblasts. METHODS: Human nasal mucosa fibroblasts were primarily cultured, and exposed to various concentrations of MMC and 5-FU for 5 minutes. Control fibroblasts were exposed to only DMEM media without the drugs. Effect of drugs on cell morphology was observed by phase-contrast microscopy. Cell viability and apoptosis were measured using MTT [3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyl-tetrazolium bromide] assay and Acridine orange/Hoechst (AO/HO) staining, respectively. RESULTS: In both experimental groups exposed to MMC and 5-FU, fibroblasts maintained standard spindle shape. The MTT assay showed that both MMC and 5-FU inhibited fibroblast proliferation in a dose dependent manner. AO/HO staining showed apoptotic cells in both experimental groups. CONCLUSIONS: Both MMC and 5-FU have an antiproliferative effect on fibroblasts in vitro at least through induction of apoptosis. Therefore, adjuvant use of either MMC or 5-FU during endonasal dacryocystorhinostomy may improve the clinical outcome by inhibiting proliferation of the nasal mucosa.
Antimetabolites ; Apoptosis ; Cell Survival ; Dacryocystorhinostomy ; Fibroblasts ; Fluorouracil ; Humans ; Microscopy, Phase-Contrast ; Mitomycin ; Nasal Mucosa

Differentiation of renal(RH) and non-renal(NRH) hematuria is important in the diagnosis and treatment of the patients with hematuria. Recently, urine RBC immunoperoxidase(IPx) staining method was developed, but there was no report on the usefulness of IPx in Korea. We validated the usefulness of IPx by comparing with the PCM. Both PCM and IPx were performed at the same time in 26 patients with RH confirmed by renal biopsy and 23 patients with NRH confirmed by radiologic and/or pathologic studies who were admitted to Chungbuk National University Hospital from January 1996 to December 1996. The age of RH and NRH group were 36.6+/-15.0 and 56.5+/-22.2 years. 35.7+/-30.4% of urine RBC were stained by IPx in RH group and only 1.6+/-4.4% were stained in NRH group(P<0.001). 23.4+/-29.9% of urine RBC by PCM were counted as dysmorphic RBC in RH group and 5.7+/-13.6% were counted in NRH group(P<0.05). At the cut-off value of 20%, the sensitivity and specificity of IPx were 57.7% and 100%. At the cut-off value of 30%, those of PCM were 30.9% and 95.7%, respectively. When comparing overall test performance by calculating AUCs of ROC(receiver operating characteristics) curve, IPx was better than PCM. IPx was better than PCM in localizing the origin of hematuria. The NRH might be excluded when IPx(+) cells are more than 20% of total urine RBC.
Area Under Curve ; Biopsy ; Chungcheongbuk-do ; Diagnosis ; Hematuria* ; Humans ; Korea ; Microscopy, Phase-Contrast* ; Sensitivity and Specificity ; Uromodulin