• Title/Summary/Keyword: Fixative

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Determining the Proportions of Bone and Cartilage Growth in the Crucian Carp (carassius auratus) Using the Modified Simultaneous Differential Staining Technique

  • Lee, Jin-Heon
    • 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.

Improved Methodology for Identification of Cryptomonads: Combining Light Microscopy and PCR Amplification

  • Xia, Shuang;Cheng, Yingyin;Zhu, Huan;Liu, Guoxiang;Hu, Zhengyu
    • Journal of Microbiology and Biotechnology
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    • v.23 no.3
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    • pp.289-296
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    • 2013
  • Cryptomonads are unicellular, biflagellate algae. Generally, cryptomonad cells cannot be preserved well because of their fragile nature, and an improved methodology should be developed to identify cryptomonads from natural habitats. In this study, we tried using several cytological fixatives, including glutaraldehyde, formaldehyde, and their combinations to preserve field samples collected from various waters, and the currently used fixative, Lugol's solution was tested for comparison. Results showed that among the fixatives tested, glutaraldehyde preserved the samples best, and the optimal concentration of glutaraldehyde was 2%. The cell morphology was well preserved by glutaraldehyde. Cells kept their original color, volume, and shape, and important taxonomic features such as furrow/gullet complex, ejectosomes, as well as flagella could be observed clearly, whereas these organelles frequently disappeared in Lugol's solution preserved samples. The osmotic adjustments and buffers tested could not preserve cell density significantly higher. Statistical calculation showed the cell density in the samples preserved by 2% glutaraldehyde remained stable after 43 days of the fixation procedure. In addition, DNA was extracted from glutaraldehyde preserved samples by grinding with liquid nitrogen and the 18S rDNA sequence was amplified by PCR. The sequence was virtually identical to the reference sequence, and phylogenetic analyses showed very close relationship between it and sequences from the same organism. To sum up, the present study demonstrated that 2% unbuffered glutaraldehyde, without osmotic adjustments, can preserve cryptomonads cells for identification, in terms of both light microscopy and phylogenetic analyses based on DNA sequences.

Experimental Study on Poultices Applying to Remove Fixative (Paraloid B72) on Earthen Mural Painting

  • Lee, Kyeong Min;Moon, Hye Young;Yu, Yeong Gyeong;Kim, Soon Kwan
    • Journal of Conservation Science
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    • v.34 no.6
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    • pp.569-580
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    • 2018
  • The possibility of applying poulticing was studied for removing Paraloid B72, a resin used for fixing an earthen mural painting. Five types of poultices were selected from clay and gel types, and acetone, ethanol, and methyl ethyl ketone(MEK) were used as mixed solvents. The possibility of mixing between the poultice and solvent was investigated, and then the spreadability, fluidity, acidity, drying properties, and solubility of the poultices were examined to confirm the characteristics. A poultice agent, which is suitable for applying to a mural painting, was selected and applied to a painting layer sample coated with Paraloid B72. As a result, all painting layers were good condition at under 50% of the solvent. The removal efficiency of Paraloid B72 was more effective when most of the solvents were used at 50%, rather than at 25%. However, it is difficult to mix 50% of MEK and ethanol with the gel-type poultices. When used at 25%, the removal performance was poorer than that at 50%, but the mixing with all the poultices was successful. In addition, the adsorption and removal power of the gel-type poultice were better than those of the clay-type, but the latter was expected to be more suitable in short-time cases due to its high drying speed. The results of this study show that the dissolution performance varies depending on the poultices and solvents. Hence, the poultice and solvent should be selectively applied, considering the fixative of the mural to be removed.

Localization of Glutamate-immunoreactive Neural Elements in the Dog Basilar Pons

  • Lee, hyun-Sook
    • Animal cells and systems
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    • v.1 no.2
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    • pp.381-388
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    • 1997
  • Glutamate is a putative excitatory neurotransmitter in the central nervous system. The present study utilizing monoclonal antibodies against fixative-modified glutamate analyzed the distribution of glutamate-immunoreactive neuronal elements in the dog basilar pons. The glutamatergic neurons were present throughout the rostrocaudal extent of the basilar pons, predominantly to the medial and ventral subdivisions. Labelled cells were relatively sparse in the midline region of the medial nucleus and most lateral area of the lateral nucleus. The majority of glutamate-immunoreactive neuronal somata in the basilar pons was multipolar-shaped, and the size was in the range of 15-25 ${\mu}$m in diameter. Glutamate-immunoreactive axons and terminals were also observed at specific regions of the basilar pons. These observations provide evidence that this excitatory neural element functions in a multisynaptic pathway involving glutamatergic afferents to the basilar pons, pontocerebellar projection neurons, and the granule cells of the cerebellar cortex.

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Studies on Nitrosamine(I) - Contents of Nitrite and Nitrate in Korean Foods(Kimchi) and Human Saliva - (Nitrosamine에 관(關)한 연구(硏究)(제(第)1보(報)) - 한국식품(韓國食品)(김치류(類)) 및 인타액중(人唾液中) 아초산근(亞醋酸根), 초산근등(醋酸根等)의 함량(含量)에 대(對)하여 -)

  • Kwon, Hyuk-Hee
    • Journal of Nutrition and Health
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    • v.7 no.4
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    • pp.21-23
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    • 1974
  • Nitrite, one of the precursors of dimethylnitrosamine distributes widely in natural foods and also used as a color fixative in meat products. In this paper Nitrites and Nitrates contents in Korean-Kimchi and Human Saliva were as follows. (1) pH: $pH:\;3.8{\sim}5.8$(in Korean-Kimchi soup) (2) NaCl: $1.5{\sim}3.1%$(in Korean-Kimchi soup) (3) Acidity: $0.38{\sim}0.62%$(in Korean-Kimchi soup) (4) $NO_2-$ $0.25{\sim}0.68ppm$ (in Korean-Kimchi) $1.9{\sim}5.0ppm$ (in Human Saliva) (5) $NO_3-$ $35.0{\sim}92.0ppm$ (in Korean-Kimchi) $7.6{\sim}28.0ppm$ (in Human Saliva) These results showed low contents in Korean-Kimchi and Human Saliva.

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Studies on Distribution of Secondary Amines in Raw Marine Fishes (해산류(海産類)중의 제(第)2급(級) 아민분포(分布)에 관(關)한 연구(硏究))

  • Kim, Kwang-Ho;Oh, Young-Bok
    • Journal of Nutrition and Health
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    • v.11 no.1
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    • pp.17-20
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    • 1978
  • The materials carcinogenic agent, nitrosoamine, is distributed in food circumstances, and is formed when both nitrite and secondary amine are present. Nitrites are added to fishes as a color fixative or preservative and secondary amines exist in fishes. In order to find the distribution and contents of secondary amines, analysis of 19 kinds of fish was conducted. The results showed that, significant differences were not observed in the quantity of secondary amines in 19 kinds of fish by district. The highest value was 20.29 ppm in Theragra chalocogramma and the lowest was 0.022 ppm in Ostrea denselamellosa. Six fishes below 1 ppm were Nibea imbricata, Misgurnus mizolepis, Astroconger myriaster, Evynnis Japonica, Ostrea denselamellosa and Solen gouldi. Dimethyl amine only was identified and diethylamine or diphenylamine was not detected in all kinds of 19 fishes.

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A Study on the Methodology of Chromosome Preparation from Blood Culture (혈액세포를 이용한 염색체 분리 분석에 관한 방법적 고찰)

  • Sohn, S.H.;Chung, K.M.
    • Clinical and Experimental Reproductive Medicine
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    • v.21 no.2
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    • pp.207-214
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    • 1994
  • This study was carried out to develop the methodology of chromosome preparation from blood cultures in mammals which included human, mouse, cattle and pig. For karyotyping, 0.5-5.0ml of peripheral blood were collected and cultured. The satisfactory results were obtained from macroculture and microculture in all species. In culture, the patterns of cell growth were no difference among media except serum concentration and mitogen supplement. The presence of mitogen and fetal bovine serum in medium significantly affected the mitotic index. The optimal culture condition was 37$^{\circ}C$ for 3 days. And the concentration of colcemid and reincubation time also affected the chromosome morphology. In harvest, chromosome patterns were mainly affected on hypotonic treatment which included treated time and temperature, dropwise of fixative solution, and drying after slide preparation.

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고분자 물질을 사용한 토양 중 방사성 핵종의 고정화

  • 원휘준;김계남;김민길;오원진
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2003.09a
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    • pp.293-297
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    • 2003
  • A series of laboratory experiments was conducted to investigate the fixation characteristics of poly(vinylalcohol)-poly(methacrylic acid)(PVA-PMAA) mixed solution on the soluble (equation omitted)-radionuclides. Using the potentiometric titration technique, it was found out that the PVA and PMAA in a solution form intermacromolecular complex. The mobilized portion of each radionuclide by water from sand surface treated with a fixative was measured by ${\gamma}$-ray spectroscopy, The mobilized portion of minor radionuclides such as $^{241}$ Am, $^{154}$ Eu, $^{155}$ Eu and $^{144}$ Ce were higher than those of $^{134}$ Cs and $^{137}$ Cs. The capability of PVA-PMAA system was better among the candidate solutions for the fixation of total (equation omitted)-radioactivity, $^{134,137}$Cs which is composed of more than 85 % of total ${\gamma}$-radioactivity could be fixed effectively by the PVA-PMAA solution.

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The BASF Laser Optical Pitch Particle Counter - Current Applications and Future Developments

  • Champ Simon;Hughes David;Lee Woo-Suk;Esser Anton;Kaub Hans Peter
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 2006.06a
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    • pp.73-78
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    • 2006
  • The occurrence of detrimental substances in the paper machine leads to reduced economic performance of paper mills through reduced capacities due to increased paper breaks and cleaning times. In this article the latest generation of BASF's laser optical pitch particle counter is employed in the examination of a paper mill operating two paper machines and using a mixture of thermo-mechanical pulp and recycled fibres. The potential of the new instrument to differentiate between particles on the basis of size and chemical composition is discussed. In addition, direct evidence of the periodic and locational variation of detrimental substances in the paper machine is given. Finally, an example of the application of BASF's laser optical pitch particle counter in the selection of a suitable fixative system for a complex furnish is presented.

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Salvage rapid maxillary expansion for the relapse of maxillary transverse expansion after Le Fort I with parasagittal osteotomy

  • Lee, Hyun-Woo;Kim, Su-Jung;Kwon, Yong-Dae
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.41 no.2
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    • pp.97-101
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    • 2015
  • Maxillary transverse deficiency is one of the most common deformities among occlusal discrepancies. Typical surgical methods are segmental Le Fort I osteotomy and surgically-assisted rapid maxillary expansion (SARME). This patient underwent a parasagittal split with a Le Fort I osteotomy to correct transverse maxillary deficiency. During follow-up, early transverse relapse occurred and rapid maxillary expansion (RME) application with removal of the fixative plate on the constricted side was able to regain the dimension again. RME application may be appropriate salvage therapy for such a case.