• Title/Summary/Keyword: microwave fixation

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Fixation of Cellular Ultrastructure by the Microwave Irradiation (마이크로파 조사에 의한 세포 미세구조의 고정 효과)

  • Shin, Kil-Sang;Kim, Wan-Jong;Jeon, Jin-Seok
    • Applied Microscopy
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    • v.26 no.4
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    • pp.401-410
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    • 1996
  • The microwave fixator has recently been introduced in morphological research. The present study was carried out to investigate the ultrastructural effects of microwave fixation of rat brain. kidney, liver and skeletal muscle tissues. The results are as follows: In the case of microwave fixed cerebrum. the cytoplasmic processes of neurons and the various membranous organelles such as nuclear envelope, mitochondria, rough endoplasmic reticulum and Golgi apparatus were well preserved, The myelin sheath wrapping neuronal axon was prominent. Microwave fixed hepatocytes showed the microvilli on the free surface of bile canaliculus, the evident nucleolar components, and typical organelles. In nephron, ultrastructures of glomerulus and Bowman's capsule were preserved, and also tubular wall were structurally observed. Among the skeletal muscle cells, plentiful collagen fibers were appeared, myofibrils and mitochondria were typically observed. In conclusion, the microwave fixation procedures result in an good preservation of the tissues and would be time- and reagent-saving.

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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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Microwave fixation of Setaria Digitata for scanning electron microscopy (선충류의 주사전자현미경적 관찰을 위한 마이크로웨이브 고정법)

  • Lee, San-Soo;Cho, Kyoung-Oh;Shin, Kil-Sang;Shin, Sung-Shik
    • Korean Journal of Veterinary Research
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    • v.47 no.2
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    • pp.203-207
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    • 2007
  • Conventional processing of biological materials including nematode parasites for scanning electron microscopy includes fixation with glutaraldehyde and osmium, followed by dehydration in an ascending grade of ethanol, and finally freeze drying. This procedure takes about 8 to 12 h depending on the characteristics of samples. Microwave irradiation of 2,450 MHz enhance the action of cross-linking fixatives and can greatly accelerate various stages of tissue processing. In this study, samples of nematode parasites, Setaria digitata, were fixed by a combination of conventional chemical fixation and the microwave irradiation during the process. The microwave irradiation was also incorporated in the serial dehydration process with ethanol. The complete procedure from the initial fixation to the completion of dehydration with ethanol was reduced to 1 h with good preservation of the ultrastructural details of the specimens.

Effects of histochemical staining in microwave-irradiated tissues (마이크로파 처리 고정 조직의 조직염색 효과)

  • Lee, Yoon-Jin;Lee, Sang-Han
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.8
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    • pp.417-424
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    • 2019
  • Despite its superior ability to show distinct cellular morphology and for long-term storage, conventional tissue fixation by formalin has many drawback, including slower fixation, the exposure to harmful chemicals and extensive protein modification. Herein, we assessed the effects of rapid microwave-assisted tissue fixation on histological examination and on protein integrity by comparing these microwave irradiation fixated tissues with the formalin-fixed tissues. One of the paired mouse tissues (liver and kidney) was fixed in formalin and the other was fixed by using microwave irradiation in phosphate buffered saline. Each slide from the paraffin-embedded tissues was examined by H & E staining for the adequacy of fixation and by immunohistochemical staining for antigenicity in a blinded fashion. Evaluation of protein recovery and the protein quality from the fixed tissues were analyzed by the BCA method and Western blotting, respectively. The results from H & E staining and immunohistochemical staining showed that the sections obtained from microwave-fixed tissues under our experimental conditions were comparable to those of the formalin-fixed tissues except for the integrity of RBCs. Furthermore, proteins were effectively extracted from the microwave-fixed tissues with acceptable preservation of the proteins' quality. Taken together, this microwave-assisted tissue processing yields a quick fixation and better protein recovery in higher amounts, as well as the adequacy of fixation and the antigenicity being comparable to formalin-fixed tissues, and this all suggests that this new fixation technique can be applied in an environment where rapid tissue fixation is required.

The Fixation Effects in Immunohistochemistry and Electron Microscopy Using Low Energy of Microwave (LEM) in Human Gastric Adenocarcinoma and HeLa Cell (사람 위선암과 HeLa 세포에 관한 저에너지 마이크로파 고정효과의 조직화학 및 전자현미경적 연구)

  • Yang, Seung-Ha;Son, Tae-Ho;Shin, Kil-Sang
    • Applied Microscopy
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    • v.31 no.2
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    • pp.185-197
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    • 2001
  • Human gastric adenocarcinomas are fixated with low energy of microwave (LEM) to study fixation effects in level of ultrastructure and antigenicity of the cancer. For the Ag-Ab reactions , the LEM fixated sdenocarcinomas are incorporated with monoclonal mouse anti-human p53 (IgG2b, kappa) and rabbit anti human cerbB-2. The retrieval of antigenicity are easily recognizable in the LEM fixated sections compared with that of frozen sections which show often diffused colour reactions. And the LEM fixation methods have preserved ultrastructures of the adenocarcinoma, but it was often difficult to maintain constancy in fixation effects. For the constancy, LEM was coupled with low concentration of chemical fixatives, such as glutaraldehyde (<1%) and $OsO_4$ (<0.5%). The results were acceptable, but there are tendencies that the adenocarcinoma requisitioned rather weak microwave energy to come into the optimal fixation effects. Therefore , cultured HeLa cells were fixated with lower energy of microwave than that used to the adenocarcinoma. The ultrastructures of the single HeLa cell have been preserved. The results may imply that a different energy levels of microwave are requisitioned in accordance with kinds of cells and tissues for the optimal fixation effects. It is reported and discussed that the fixation methods of LEM used in this work could be applied routinely to conceal a insufficient diffusion rate of chemical fixatives into some kinds of cancer without compromising the ultrastructures as well as to improve antigenic quality of frozen sections.

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Fixation and Histochemistry of Biological Tissues Using the Microwave Fixator Equipped with Infrared-Temperature Sensor (적외선 온도감응기를 장착한 마이크로파 고정기에 의한 생체조직 고정효과와 조직화학적 특성)

  • 신길상;민소연;김완종;손태호
    • The Korean Journal of Zoology
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    • v.38 no.3
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    • pp.417-425
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    • 1995
  • The present study was carried out to investigate the effect of microwave fixation in comparison with that of chemical fixation in preparing the microscopic samples. The microwave fixator was equipped with infrared-temperature sensor, and that was designed to compensate air temperature in the microwave fixator. In the microwave fixation, rat tongue was well preserved in terms of muscular fasciculus and pancreas stained by Feulgen reagents showed clear reaction products in the nucleus. Reaction products by PAS method in duodenal villi appeared specifically at the goblet cells. In electron microscopy, pancreatic cellular components such as secretory granules and collagen bundles were well preserved in both fixations. In aspect of histochemical reaction and electron microscopy, high quality was due to the protein content of microwave fixed specimen. The microwave fixation method saved total duration engaging microscopic preparation.

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A Study on the Dyeing of Polyester Fabric by Microwave Heating(I) (Microwave 가열에 의한 Polyester직물의 염색에 관한 연구(I))

  • 서수정;임수경;김삼수
    • Textile Coloration and Finishing
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    • v.10 no.4
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    • pp.7-14
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    • 1998
  • Microwaves are high frequency radiation capable of generating very rapid, uniform and efficient heating of textile material. Microwave heated dyeing of polyester fabric was tried with different solvent systems, irradiation time and dye concentration. Microwave fixation methods were used with 100% water,30% urea, EG and DMF, respectively, pad-baths in which the padded fabrics were exposed over a heated or boiling water bath to maintain sufficient moisture content during irradiation. In order to ascertain the relation between the dyeing property of polyester fabric and the microwave irradiation condition caused by microwave heating, the K/S values and fastness properties of dyed fabrics such as light, washing and sublimation fastness were measured.

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The Fixation Effects of Biological specimen Using Microwave Oven Equipped With Infrared-Temperature Sensor (적외선 온도감응기를 장착한 마이크로파 오븐의 생체물질 고정효과)

  • 신길상
    • The Korean Journal of Zoology
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    • v.37 no.2
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    • pp.144-155
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    • 1994
  • 마이크로파 에너지로서 현미경 관찰을 위한 생체물질을 고정할 때, 고정효과는 화학 고정액을 사용할 때와 비교하여 보다 우수한 또는 적어도 동일한 고정효과를 볼 수 있다는 것은 알려진 사실이다. 그러나 기존의 마이크로파 오븐을 사용하여 생체물질을 고정하면 고정 정도에 일정성이 없는데, 이는 마이크로파가 균일하게 조사되지 않고 고정에 적절하도록 마이크로팍의 강도를 조절하기 어려우며, 이로 인하여 정확한 고정온도의 측정이 불가능하기 때문으로 생각된다. 이 연구에서는 마이크로파가 조사되는 동안 시료의 온도를 측정할 수 없는 기존의 마이크연.파 오븐의 단점을 보완하기 위하여 비접촉식 적외선 온도 감응기를 장착한 마이크로파 오븐을 개발하였으며 고정효과를 관찰하기 위하여는 포유류의 기준 조직으로 알려진 생쥐의 신장을 고정하였다 고정효과는 전체적인 구조와 세부 구조의 보존이 모두 우수하였고 최적의 고정온도는 28$\pm$1$^{\circ}C$(완충용액 20 ml, 10초) 이었다. 이와같이 고정된 재료는 고정효과 뿐만 아니라 염색성이 좋아서 조직화학 반응이 우수하고, 냉동절편법과 연계했을 때 전체 절편제작 과정이 2~4시간 내에 완료될 수 있었다 이때 절편의 질(질)은 화학 고정액이나 냉동절편 법에 의하여 제작된 것 보다 우수하였다.

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Application on Microwave Energy in the Preparation of Fish Samples for Electron Microscopic Observation

  • Kim Soo Jin;Oh Hae Keun;Song Young-Hwan;Chung Hyun-Do;Kim Young-Tae;Park Nam-Kyu;Choi Tae-Jin
    • Fisheries and Aquatic Sciences
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    • v.1 no.2
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    • pp.187-191
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    • 1998
  • Chemotherapy can not be applied for the control of fish viral diseases because viruses depend on host machinery for their replication. Although new control strategies including vaccination are under development, avoidance of virus introduction by rapid and correct diagnosis is the best way of fish viral disease control. Although observation of virus particles with an electron microscope is an easy method for virus detection, it take a few days for the sample preparation. In order to shorten the sample preparation time, microwave radiation was applied in the procedure. With this method, 15 seconds was enough for fixation of virus infected fish samples or cultured cells inoculated with infectious hematopoietic necrosis virus, which takes 2-4 hours with routine methods. Also four minutes was enough for polymerization of embedding resin which takes 24-48 hours with routine methods. Samples prepared with microwave were good enough for direct electron microscopic observation and immunogold labeling assay.

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