• Title/Summary/Keyword: chemical fixation

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Ultrastructures of Ptilota filicina (Rhodophyta) by High Pressure Freezing(HPF): Comparison of HPF Fixation and Chemical Fixation (High Pressure Freezing (HPF)을 이용한 조류 Ptilota filicina의 미세구조 관찰:HPF 고정법과 화학 고정법의 비교)

  • Lee, Sang-Hee;Kim, Youn-Joong;Jeong, Jong-Man;Kim, Jin-Gyu;Kim, Young-Min;Kweon, Hee-Seok;Moon, Won-Jin;Lee, Seok-Hoon
    • ALGAE
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    • v.21 no.4
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    • pp.479-483
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    • 2006
  • In preparation of the biological samples for electron microscopy, the chemical fixation by glutaraldehyde, paraformaldehyde, and OsO4 has been generally used for a long time. However, the chemical fixation method has some problems: the infiltration time is a little bit long and the ultrastructure of cell or tissue transforms before complete fixation of sample. So, recently, cryo-fixation is considered more often in biomedical field. In this study, we compared High Pressure Freezing (HPF) method with chemical fixation method using a algal sample (Ptilota filicina J. Agardh), which was difficult to fix using chemical fixation method. In chloroplast, the ultrastructure of thylakoid lamella and phycobilisome can not show clearly by chemical fixation. In this study we could observe the ultrastructure of thylakoid lamella and phycobilisome of chloroplast very clearly using HPF fixation. An improved images of ultrastructures of nucleus, mitochondrion and floridean starch could obtain. These results suggest that HPF method is very useful method in algal specimen for electron microscopy.

Enhancement of Carbon Dioxide Fixation by Alteration of Illumination during Chlorella Vulgaris-Buitenzorg's Growth

  • Wijanarko Anondho;Dianursanti Dianursanti;Gozan Misri;Andika Sang Made Krisna;Widiastuti Paramita;Hermansyah Heri;Witarto Arief Budi;Asami Kazuhiro;Soemantojo Roekmijati Widaningroem;Ohtaguchi Kazuhisa;Koo Song-Seung
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.6
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    • pp.484-488
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    • 2006
  • Alteration of illumination with optimum carbon dioxide fixation-based curve in this research successfully enhanced the $CO_{2}-fixation\;(q_CO_{2}$ capability of Chlorella vulgaris Buitenzorg cultivated in a bubble column photo bioreactor. The level of $CO_{2}$ fixation was up to 1.91 times that observed from cultivation with intensification of illumination on an optimum growth-based curve. During 144 h of cultivation, alteration of light intensity on an optimum $CO_{2}-fixation-based$ curve produced a $q_CO_{2}$ of $12.8\;h^{-1}$. Meanwhile, alteration of light intensity with a growth-based curve only produced a $q_CO_{2}$ of $6.68\;h^{-1}$. Increases in light intensity based on a curve of optimum $CO_{2}-fixation$ produced a final cell concentration of about 5.78 g/L. Both cultivation methods were carried out under ambient pressure at a temperature of $29^{\circ}C$ with a superficial gas velocity of $2.4\;m/h(U_{G}$. Cells were grown on Beneck medium in a 1.0 L Bubble Column Photo bioreactor illuminated by a Phillips Halogen Lamp (20 W/12 V/50 Hz). The inlet gas had a carbon dioxide content of 10%.

Improvement of Histopathological Sample Preparation by Employing Microwave Heating Method on Frozen Section Specimens

  • Ahn, Seung-Ju
    • Biomedical Science Letters
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    • v.13 no.4
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    • pp.361-368
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    • 2007
  • Biological samples can be fixed either by chemical method by using chemical solution or physical methods by using heat treatment. The problem in traditional heat fixation is unsatisfactory quality due to uneven heat conduction in specimen and loss of inner cell contents. Chemical fixation method also bears several intrinsic problems like the limit in specimen size, time consumption in fixative impregnation, and loss of low molecular weight cell components. These factors deteriorate the quality of fixed specimen, thus limit the magnification and contrast of tissue pictures. Microwave heat has been reported to be a good alternative to current chemical methods to overcome these problem. In this study, we tried to introduce the microwave energy method to routine fixation work in hospital. We replaced chemical fixative with saline to provide moderate reaction condition, and used frozen section to reduce time for sample preparation. Temperature was measured at each experiment. The fixation of rat kidney tissue with 2.45 GHz electromagnetic wave and saline showed similar result to the control group fixed with traditional chemical method. Human tumor tissue fixed with 2.45 GHz electromagnetic in frozen section was improved in terms of histochemistry of PAS and immunohistochemistry of tumor marker like cytokeratin. Total turnaround time was reduced from $24\sim38$ h to to $2\sim4$ h. In conclusion, the quality of samples prepared by microwave heating method was at least as good as that of traditional method. If the condition for the fixation of different specimens is standardized, this new method could be applied to routine work in hospital, and could save working time as well.

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Chemical Fixation of Polyelectrolyte Multilayers on Polymer Substrates

  • Tuong, Son Duy;Lee, Hee-Kyung;Kim, Hong-Doo
    • Macromolecular Research
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    • v.16 no.4
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    • pp.373-378
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    • 2008
  • A simple chemical fixation method for the fabrication of layer-by-layer (LbL) polyelectrolyte multilayer (PEM) has been developed to create a large area, highly uniform film for various applications. PEM of weak poly-electrolytes, i.e., polyallylamine hydrogen chloride (PAH) and poly(acrylic acid)(PAA), was assembled on polymer substrates such as poly(methyl methacrylate)(PMMA) and polycarbonate (PC). In the case of a weak polyelectrolyte, the fabricated thin film thickness of the polyelectrolyte multilayers was strongly dependent on the pH of the processing solution, which enabled the film thickness or optical properties to be controlled. On the other hand, the environmental stability for device application was poor. In this study, we utilized the chemical fixation method using glutaraldehyde (GA)-amine reaction in order to stabilize the polyelectrolyte multilayers. By simple treatment of GA on the PEM film, the inherent morphology was fixed and the adhesion and mechanical strength were improved. Both surface tension and FT-IR measurements supported the chemical cross-linking reaction. The surface property of the polyelectrolyte films was altered and converted from hydrophilic to hydrophobic by chemical modification. The possible application to antireflection coating on PMMA and PC was demonstrated.

Methanol fixation for scanning electron microscopy of plants

  • Ki Woo Kim
    • Applied Microscopy
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    • v.50
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    • pp.10.1-10.6
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    • 2020
  • Plant specimens for scanning electron microscopy (SEM) are commonly treated using standard protocols. Conventional fixatives consist of toxic chemicals such as glutaraldehyde, paraformaldehyde, and osmium tetroxide. In 1996, methanol fixation was reported as a rapid alternative to the standard protocols. If specimens are immersed in methanol for 30 s or longer and critical-point dried, they appear to be comparable in preservation quality to those treated with the chemical fixatives. A modified version that consists of methanol fixation and ethanol dehydration was effective at preserving the tissue morphology and dimensions. These solvent-based fixation and dehydration protocols are regarded as rapid and simple alternatives to standard protocols for SEM of plants.

Improved Ultrastructural Preservation of Retinal Cells in Drosophila melanogaster (초고압동결장치를 이용한 초파리 레티나 세포의 향상된 미세구조)

  • Mun, Ji-Young;Park, Se-Jin;Han, Sung-Sik
    • Applied Microscopy
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    • v.37 no.3
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    • pp.175-183
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    • 2007
  • The Drosophila retinal cell is widely used to study cell development and cell signaling processes. In the past decades, conventional chemical fixation had been used to study the structure of retinal cells in Droscphila. Rapid freezing methods are superior to chemical fixation methods due to their fixation speed. Some Drosophila tissues, such as the eyes, should not be freezed due to their surrounding cuticle layer. Therefore, in the case of the Drosophila retina, the benefits of high pressure freezing and freeze substitution (HPF-FS) had not been verified. In this study, a retinal cell from Drosophila melanogaster had been studied by using the HPF-FS method. Compared to chemical fixation, the preservation of the cytoplasm in the HPF-FS sample was improved on the whole. The HPF-FS cell membranes were smoother than that of chemical fixation. In addition, HPF-FS preserved the mitochondria structures very well. These results of the present study suggest that HPF-FS is superior to other fixation methods for the preservation of the retinal cell structure.

The Characteristics of Carbon Dioxide Fixation by Chlorella sp. HA-1 in Semi-continuous Operation (반연속식 운전에서 Chlorella sp. HA-1의 이산화탄소 고정화 특성)

  • Lee, Jae-Young;Kang, Hyun-Ah;Yang, Ji-Won
    • KSBB Journal
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    • v.14 no.6
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    • pp.742-746
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    • 1999
  • The microalgal, Chlorella sp. HA-1, had good $CO_2$ fixation efficiency compared to other algal strains at the same operating condition. In this study, Chorella sp. HA-1 showed similar tolerance both 10% and 20% $CO_2$ concentration. By optimization of the major operation variables such as pH, initial cell concentration, light intensity, the $CO_2$ fixation rate could be raised to a reasonably high value, 372 $gCO_2/m^2{\cdot}day$ in a 3 L internally illuminated photobioreactor. In order to maintain the $CO_2$ fixation rate for a long time, the method of semi-continuous operation was employed, in which dilution ratio was the controlling parameter. Starting with the dilution ratio of 0.5 with the increased increment of 0.1, the constant $CO_2$ fixation rate was obtained.

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Biological Fixation of $CO_2$ by Chlorella sp. HA-1 in a Semi-Continuous and Series Reactor System

  • LEE JAE-YOUNG;KWON TAE-SOON;BAEK KITAE;YANG JI-WON
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.461-465
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    • 2005
  • Characteristics of biological $CO_2$ fixation by Chlorella sp. HA-1 were investigated in a semi-continuous and series reactor system using an internally illuminated photobioreactor to overcome shortcomings of physicochemical technologies such as adsorption and membrane separation. High $CO_2$ fixation rate was achieved in the semi-continuous reactor system, in which the dilution ratios of the culture medium were controlled. The average $CO_2$ fixation rate was maintained almost constantly when the dilution ratio increased by 0.1 increment from the initial value of 0.5. The total removal efficiency of $CO_2$ was enhanced by employing a series reactor system. The average $CO_2$ fixation rate increased until 4.013 g $CO_2\;day^{-1}$ in a series operation of four reactors, compared to 0.986 g $CO_2\;day^{-1}$ in a batch operation mode. The total $CO_2$ fixation rate was proportional to the number of reactors used in the series reactor system. In the series reactor system of semi-continuous operation, a large amount of $CO_2$ was removed continuously for 30 days. These results showed that the present reactor systems are efficient and economically feasible for a biological $CO_2$ fixation.