• Korean Journal of Veterinary Research
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• v.35 no.2
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• pp.383-390
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• 1995

This study was performed to investigate the characteristics of tear staining syndrome in poodle dogs. Schirmer tear test, fluorescein dye test and measurement of the angle of bend between vertical and horizontal bony nasolacrimal duct were conducted in both poodles and German shepherd dogs. There were no significant differences between normal and tear-stained poodles in tear formation determined by Schimer tear test. However, there was significantly higher tear production in German shepherds than that in normal poodles(p<0.05). In the fluorescein dye test for the measurement of tear excretion, the dye was observed within $14.5{\pm}6.5$ minutes after dropping of the dye in normal poodles, but was not observed even over 30 minutes in tear-stained poodles. German shepherds had rather rapid passage time($0.4{\pm}0.3$ minutes) than poodles in the dye excretion. In the measurement of the angle of bend between vertical and horizontal bony nasolacrimal duct through dacryocystorhinography, there were no significant differences between normal tear-stained poodles with showing $85.0{\pm}6.8^{\circ}$ and $89.8{\pm}6.5^{\circ}$, respectively. However, obtuse angle of bend($106.8{\pm}4.7^{\circ}$) was shown in German shepherds. These results have ascertained that tear staining syndrome of poodle dogs was not related to tear production but to the rate of tear excretion.

• Korean Journal of Veterinary Research
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• v.35 no.2
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• pp.317-326
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• 1995

The purpose of this study was to establish a method for in vitro culture of C parvum isolated in Korea by determination of suitable cell model to complete development of this parasite. The result obtained were summerized as follows: 1. To determine the most suitable cell line, six types of cell line were examined by microscopy. All cell lines were infected with C parvum and showed the highest infection score in HmLu cells. 2. The staining methods including DMSO-modified acid-fast(A-F) stain, hematoxylin-eosin(H & E) stain and immunofluorescence antibody(IFA) stain were applied to examine the infection of C parvum in cell culture. These staining methods were possible to examine the infection of C parvum in cell culture. The most sensitive one was IFA staining technique. 3. Developmental stages of C parvum in HmLu cell were observed. After the initial 8 hour incubation period, some trophozoites were observed. The meronts and gametes were appeared at 24-48 hour post inoculation(PI), and oocysts were observed firstly at 48-72 hour PI. 4. In H & E stain, the parasite appeared as basophilic within parasitophorous vacuole membrane(PVM) and lying in cytoplasm at near the nucleus of the host cells. It was able to distinguish the type I, type II meronts and gametes. 5. In DMSO-modified acid-fast stain, specific stained parasites were appeared firstly after 48 hour PI. The parasites were showed with different degrees of staining bright red color within PVM. 6. The endogenous stages of parasites in HmLu cell recovered at 48, 96, 120 and 144 hour after inoculation were reacted with rabbit immunized serum in immunofluorescence antibody and avidin-biotin complex peroxidase staining technique.

• Journal of Environmental Health Sciences
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• v.36 no.4
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• pp.337-341
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• 2010

The modified simultaneous differential staining technique, which enables double staining of cartilage and bones, needs to be improved to prevent soft tissues from being damaged during the staining process. Key factors influencing the extent to which soft tissues are damaged include the fixative used, macerating time, potassium hydroxide concentration, incubation temperature and the removal of skin from specimens. Here we describe a protocol that enables the hardening of tissues during bleaching and maceration. We also describe a method for objectively measuring rates of cartilage and bone growth. The use of formalin as a fixative rendered soft tissues more rigid due to the resulting chemical bonds formed between proteins. Blotted specimens were immersed in 1% potassium hydroxide (KOH) and incubated at $37^{\circ}C$ for 1 day (smaller specimens) or 2-3 days (larger specimens). The 1% KOH solution was also used as the diluent solution for the subsequent immersion in a graded series of 30%, 50%, 70%, 90%, 100% glycerol solutions, a procedure that made soft tissues even more transparent and hardened. It was not necessary to remove the skin of specimens shorter than 2 cm, since the macerating solution could easily penetrate their thin skin layer and continuously remove those pigments hindering visibility. Since excessive osmosis is another factor that can damage soft tissues in the macerating process by causing the rupture of those cells not able to withstand the osmotic pressure, here it was minimized by balancing the salt concentration between the interior and exterior of cells with the addition of 0.9% sodium chloride (NaCl) in the macerating solution. Finally, to determine the proportions of cartilage and bone growth, photographs of the stained specimens were taken with a dissecting microscope and sections corresponding to the cartilage and bones were cut out from the printed pictures and weighed. Our results show that this method is suitable for the objective evaluation of bone and cartilage growth.

• Journal of the Korean Wood Science and Technology
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• v.33 no.2 s.130
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• pp.65-71
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• 2005

This study was performed to understand the interaction between Ophiostomataceae and basidiomycetes fungi during cultures, and whether the basidiomycetes fungi inhibit the growth and decolorize dark pigments of blue staining fungi. The conjoint cultivation was studied on 2% malt extract agar. The ability of basidial cultures to decolorize dark pigments of ophiostomatoid fungi was the main characteristics estimated during this study. More than half of basidial cultures were characterized by deadlock interaction with blue staining fungi. In the dual cultures, where basidial partners were presented by Agaricus bisporus(64), Laetiporus sulphureus(L01/89), Trametes versicolor(09) and unknown fungus(02), antagonism was found at the phase of primary contact of colonies. Replacement interaction resulted usually in decreasing dark colour of substrate was observed for 11 basidial cultures that were belonging mainly to white-rot fungi. Among them Abortiporus biennis(123), Antrodiella hoehnelii(S28/91), Bjerkandera fumosa (137), and Gleophyllum odoratum(124) were characterized by the absence of deadlock-phase: they began to grow over dark colonies of their partners just after primary contact. Basidiomycetes did not affect strongly the pigments of Ceratocystis spp. and Leptographium sibirica isolates, but completely decolorized colonies of Ophiostoma ips and to a smaller degree Ophiostoma minus. Antrodiella hoehnelii(S28/91), Bjerkandera fumosa(137), Gleophyllum odoratum(124) and Trametes versicolor(B18/91) cultures were found to be the most active in decreasing dark color of blue staining fungi colonies. The cultures were recommended for further development as agents of biopulping of wood chips and bio-control of blue stain in woods.

• Applied Microscopy
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• v.42 no.1
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• pp.49-52
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• 2012

For TEM study of biological samples or polymers that are contained in organic structure, it is often required that the sample is prepared by using ultramicrotome and stained with proper agents to increase the contrast of organic structure. In this study, we investigated an efficient TEM sample preparation method for ductile heterogeneity material by using ultramicrotomy. Cryo-ultramicrotomy is a suitable method that is capable of rendering sample hardness for various ductile materials. However, it has several factors to consider, such as experimental cost, working time and finding the optimal staining conditions. To satisfy these considerations, we prepared TEM sample by using ultramicrotome without cryofunction, and secured the sample hardness by applying the staining process prior to ultrathin sectioning. The cross-linked polyethylene structure in the sample was stained with the 2% $RuO_4$ solution in a sealed test tube for 24 hours at $4^{\circ}C$. After the sample staining, ultrathin sections of sample were prepared using ultramicrotome. As a result, it was revealed that the difficulties associated with staining of ultrathin sections prepared by low-temperature conditions were improved. In addition, appropriate staining depth of sample could be selected for sectioning process. The quality of TEM sample obtained by using this method was better than that of cryo-ultramicroscopy. Finally, it is expected that our method could be effectively applied in TEM sample preparation for a variety of nano-bio convergence materials.

• Journal of the korean academy of Pediatric Dentistry
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• v.23 no.2
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• pp.559-574
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• 1996

The purpose of this study was to determine whether there was any correlation between temporomandibular joint dysfunction and structure of the mandibular condyle. Weanling rats had their masseter muscles resected and immunohistochemical findings were observed with a light microscope. The results obtained were as follows : 1. The condylar cartilage region was divided into articular, proliferating, cartilage cell and hypertrophic cell layers according to cell morphology. 2. In light microscopic views, the proliferating and cartilage cell layers of the experimental group decreased gradually and at the 8th week significantly. 3. In immunohistochemical staining for type I and II collagen, a reaction was detected in the lower part of proliferating cell and cartilage cell layers. In the cartilage cell layers, a stronger cellular reaction was present. Immunohistochemical staining for type II collagen reacted more strongly than that of type I collagen. 4. In immunohistochemical staining for proteoglycan, the staining of the experimental group resembled the control group and gradually showed a weak reaction. The proliferating and cartilage cell layers reacted more strongly than the hypertrophic cell layer. 5. In immunohistochemical staining for proliferating cell nuclear antigen(PCNA), the strong reaction was detected in the nucleus of the proliferating cell layer both in control and experimental groups. But the thickness of the proliferating layer decreased in experimental group, consequently the reaction of the experimental group was reduced more than that of the control group.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.29 no.5
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• pp.405-416
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• 2007

Objective : Lots of papers have revealed that tumor growth related factors such as EGF, EGFR, c-erbB-2 play an important role in tumorigenesis and proliferation. These factors are found in most tumors of ectodermal origin. But, documentations of tumor growth related factors on salivary gland tumors were rare. Therefore, we determined expressions of tumor growth related factors; PCNA, p53, EGF, EGFR, cerbB2(HER-2), Maspin, DMBT-1, N-Ras in representative salivary gland tumors. Materials and methods : A few types of salivary tumors were examined by immunohistochemical assays. Each antibody was applied to specimens of tumors. Specimens were composed of 5 pleomorphic adenomas (PA), 3 mucoepidermoid carcinomas (MEC), 2 adenoid cystic carcinomas (ACC) and 2 squamous cell carcinomas (SCC) from 12 patients. One specimen was selected randomly as negative control. For evaluation of staining intensity, each stained sample was divided into 5 grade; no staining, obscure, weak staining, moderate staining, strong staining. Results : Strong expressions of PCNA were found in all tumors except of PA. EGF was expressed strongly in SCC, ACC sequently. But in both PA and MEC, EGF expression was weak. EGFR and c-erbB-2 expression showed similar patterns in all salivary gland tumor tissues. P53 showed weak expression generally in all salivary gland tumors. DMBT-1 was expressed in SCC rather than in ACC or in MEC. N-Ras showed weak expressions in all salivary gland tumors except of squamous cell carcinoma. Conclusion : Taken together, tumor growth related factors were expressed in salivay tumors as well as mucosal squamous cell carcinoma. Especially EGFR and c-erbB-2 could be candidates as diagnostic markers for estimating clinical grade of salivary gland tumors. But further studies with reliable methods will be needed to confirm the results of this study.

• Journal of Embryo Transfer
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• v.30 no.3
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• pp.213-218
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• 2015

In the study for a differentiation and development of spermatogonial cells, the researchers should commonly require a simple, fast and reasonable method that could evaluate the developmental stage of male germ cells without any damage and also relentlessly culture them so far as a cell stage aiming at experimental applications. For developing the efficient method to identify the stage of sperm cells, the morphological characteristics of sperm cells were investigated by staining the cells with blue fluorescent dye Hoechst 33258, and a criterion for male germ cell classification was elicited from results of the previous investigation, then the efficiency of the criterion was verified by applying it to assort the germ cells recovered from male mice in age from 6 to 35 days. As morphological characteristics, spermatogonia significantly differed from spermatocytes in size, appearance and fluorescent patches of nucleus, and spermatids could also be distinguished from spermatozoa by making a difference in the volume and shape of nucleus and the shape and fluorescence of tail. Aforesaid criterion was applicable for classifying in vitro cultured sperm cells by verifying its efficiency and propriety for assorting the stages of testicular germ cells. However, the fluorescent staining showed that germ cells in mouse testis should be dramatically differentiated and developed at 21 days and 35 days of age, which were known as times of sexual puberty and maturity in male mice, respectively. In conclusion, the results indicated that this simple criterion for sperm cell classification using fluorescence staining with Hoechst 33258 may be highly efficient and reasonable for spermatogenesis study.

• Journal of Korean Society on Water Environment
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• v.40 no.1
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• pp.36-46
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• 2024

Microplastics in water resources have been recognized as a serious problem. The discharge of microplastics from wastewater treatment plants is considered a major contributor to environmental pollution in water resources. However, a reliable analytical method for quantifying microplastics in wastewater treatment plants has not yet been established. This study proposes a reliable, quick, and easy analytical method for quantifying microplastics. For the removal of organic particles, preprocessing steps were applied including oxidation, sonication, washing, and sieving. Nile Red staining was used to visualize microplastics, and quantitative analysis was conducted using fluorescent imaging. The stained microplastics were ultimately quantified through image analysis software. Among the preprocessing steps, sonication and washing stages were particularly effective in efficiently removing interfering substances from wastewater, enhancing the accuracy of the microplastic analysis. Additionally, various solvents (methanol, acetone, and N-hexane) for the Nile Red staining solution were tested. When N-hexane was applied as the solvent, the quantity of stained microplastics was lower compared to methanol and acetone. This suggests that N-hexane has a greater potential of reducing false staining and counting of non-plastic particles. In summary, this research demonstrates a robust method for quantifying microplastics in wastewater treatment plants by employing effective preprocessing steps and optimizing the staining process with Nile Red and N-hexane.

• Korean Journal of Veterinary Research
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• v.15 no.1
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• pp.15-17
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• 1975

In order to investigate the longevity of the reticulocyte in peripheral blood, samples were taken from the jugular vein at birth; 2nd, 4th, 7th, and 14th day after birth in eight healthy Korean native goats (body weight 1.5 to 2.0kg). The samples were stained in 1, 2, 3, 4, 6, 10, 24, and 48 hours after bleeding, and observed the reduction in the number of reticulocytes and the degree of the staining. The data have shown that the number and staining degree of reticulocytes in Korean native goots were appeared as normal untill 24 hours after bleeding, however, the number and staining degree were reduced in 48 hours after bleeding.