• Title/Summary/Keyword: Glutaraldehyde

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AN ELECTRON MICROSCOPY OF SPERMIOGENESES IN Clonorchis sinnensis (간(肝)디스토마 정자완성(精子完成)의 전자현미경적(電子顯微鏡的) 연구(硏究))

  • Paik, Kyong-Ki;Lee, Uen-Ho
    • Applied Microscopy
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    • v.1 no.1
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    • pp.35-42
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    • 1969
  • 가토간장에서 채취(採取)한 간(肝)디스토마 성충(成蟲)의 정소(精巢)와 수정낭(受精囊)을 1.25% glutaraldehyde와 1% 사산화(四酸化) 오스뮴산(酸)으로 냉실(冷室)에서 이중고정(二重固定)하였다. 고정(固定)된 시료(試料)는 양식(樣式)에 따라 탈수(脫水)한 후(後) Epon-812로 포매(包埋)하여 MT-2형(型) Porter Blum microtome으로서 초박절편(超薄切片)을 만들어 수사화연(水酸化鉛)과 초산(醋酸)우라닐로서 이중염색(二重染色)한후 Hitachi HS-7형(型)과 HU-11E형(型)의 전자현미경(電子顯微鏡)으로 관찰(觀察)하였다. 관찰결과 간(肝)디스토마의 정세포(精細胞)는 타원형으로 난형(卵形)의 인(仁)을 포함(包含)한 큰 핵(核)을 갖고 있으며 핵(核)을 둘러싼 비교적소량(比較的小量)의 세포질(細胞質)에는 낭형(囊形)의 조면소포체(粗面小胞體)가 희소(稀少)하게 있으며 유리(遊離)리보좀 및 즐(櫛)을 가진 미토콘드리아, 중심체(中心體), 층판상(層板狀)의 골지체가 있다. 정자(精子)는 긴 원주형(圓柱形)의 핵(核)과 선단(先端)에 첨체(尖體)를 포함(包含)한 두부(頭部)와 중종편(中終片), 미부(尾部)의 말단부(末端部)로 갈수록 점차 가늘어져서 결국(結局) 편모(鞭毛)로 끝난다. 두부(頭部)의 선단부(先端部) 가까이 핵환(核環)이 있으므로 이를 기시점(起始點)으로 직경(直徑) $250{\AA}$ 정도의 $8{\sim}10$개(個)의 미세소관(微細小管)이 축사(軸絲)의 배면(背面) 원형질막(原形質膜) 직내면(直內面)의 외형질(外形質)에 평행(平行)하게 중종편(中終片)까지 신장(伸長)되어 있다. 미토콘드리아는 융합(融合)되어 두부(頭部)의 후반부(後半部) 핵(核)의 복측(腹側)에 평행(平行)하게 축사(軸絲)를 감싸는 원추형(圓錐形)으로 종단면(縱斷面)에서 반점상(班點狀)의 횡문(橫紋)이 관찰(觀察)되었다. 중심체(中心體)에서 기원(起原)된 축사(軸絲)는 중심부(中心部)에 중심섬유(中心纖維)와 중심초 및 이중(二重)의 미세소관(微細小管)이 9개(個) 둘러싸고 있는 구조(構造)를 하고 있다.

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Ultrastructural Study on the Substantia Nigra of the Head-Irradiated Rats (머리에 방사선 상해를 받은 흰쥐 흑색질의 미세구조)

  • Bae, Hack-Gun;Yang, Nam-Gil;Ahn, E-Tay;Ko, Jeong-Sik;Park, Kyung-Ho;Kim, Jin-Gook
    • Applied Microscopy
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    • v.22 no.2
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    • pp.30-45
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    • 1992
  • An experimental study on the acute irradiation effects on the substantia nigra of head-irradiated rats were carried out. Rats anesthetized with sodium thiopental, were exposed only on their head areas with a single dose of 3,000 rads or 6,000 rads, respectively. Radiation was produced by Mitsubishi linear accelerator at the speed of 200 rads/min. Aminals were sacrificed on 6 hours, 2 days and 6 days following irradiations. By the perfusion fixation through the heart, rats were fixed with 1% glutaraldehyde-1% paraformaldehyde solution. Two hours later, brains were exposed and immersed in the same fixatives over night. Tissue blocks from subtantia nigra were punched out, and they were refixed in the 2% osmium tetroxide solution. Blocks were dehydrated through alcohol series, and embedded in the araldite mixture. Ultrathin sections were stained with uranyl acetate and lead citrate solutions, From the ultrastructural study, following results were made: 1. Six hours after irradiation, severe depletion of synaptic vesicles was occurred in the many axon terminals of the nigral neuropil. 2. Dramatical decrease of lysosomes and dense granules was observed. 3. Two days following irradiation, alterations of ribosomes, granular endoplasmic reticula, mitochondria, etc, were noticed. 4. Many of the malformations were seen to be repaired on the 6th day. 5. Above results were interpreted as follows. At the acute stage of heavy irradiation, neurotransmitters in the substantia nigra are released severely. But they are recovered within 6 days. It is concluded that acute head-irradiation may result severe disturbance of nigral motor control function during the first few days.

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THE FINE STRUCTURE OF THE FIBROBLAST IN INFLAMED HUMAN PULP TISSUE (염증성치수조직중(炎症性齒髓組織中) 섬유아세포(纖維牙細胞)의 미세구조(微細構造)에 관(關)한 연구(硏究))

  • Bae, Kwang-Shik
    • Restorative Dentistry and Endodontics
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    • v.8 no.1
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    • pp.7-17
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    • 1982
  • The purpose of this study was to investigate the fine structural modifications of fibroblasts in the coronal region of inflamed human pulps from carious teeth. Six untreated human teeth with large carious lesions and two normal teeth as control were selected from male and female patients between the ages of 20 and 39. The teeth were divided into 4 groups by light microscopic findings: the normal control group, the chronic inflammatory cell-appeared group, the acute and chronic inflammatory cell-appeared group, and the total necrosis group. All tissues were fixed in 2.5% glutaraldehyde in 0.1M sodium cacodylate buffer at pH 7.4 and 1% osmic acid in same buffer. They were embedded in Epon 812. The ultrathin sections were stained conventionally and examined with a AEI Corynth 500 electron microscope. The results were as follows; 1. The fibroblasts of the normal pulps were almost in a quiescent state. 2. The active and the quiescent fibroblasts were found in the pulps of the chronic inflammatory cell-appeared group. Lymphocytes and plasma cells were also seen scattered among these fibroblasts. 3. In the pulps of the acute and chronic inflammatory cell-appeared group, active, degenerative and necrotic fibroblasts were found in the PMN appeared area. And all the fibroblasts in the fibrosis area were active. In the area of chronic inflammatory cellular infiltration, almost all the fibroblasts were active, but seldom were quiescent fibroblasts observed. Some fibroblasts in the pulps of two teeth had large vacuoles that contained banded collagen fibrils. The phagosomes had small beaded vesicles or large lysosome-like varicosity. In two of the teeth, microorganisms were present and two morphological shapes were identified, a rod and a coccus. 4. Vacuolar, vesicular, lamellar, fibrous and myelin structures were observed in the pulp of the total necrosis group, and cocci were also seen.

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Ultrastructural Influence of Nephrotoxic Agents on the Juxtaglomerular Cells of Mice (신장 상해물질이 생쥐 사구체곁세포의 미세구조에 미치는 영향)

  • Park, Kyung-Ho;Kim, Sang-Chul;Ahn, E-Tay;Ko, Jeong-Sik;Yang, Nam-Gil
    • Applied Microscopy
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    • v.26 no.4
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    • pp.431-446
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    • 1996
  • This experiment was performed to study the ultrastructural changes of the juxtaglomerular cell of mice following subcutaneous injection of heavy metallic agents. Male mice were divided into normal and experimental groups. The mice were subcutaneouly injected with $HgCl_2$ (2mg, 5mg or 10 mg/Kg/BW) or with $K_{2}Cr_{2}O_7$(5 mg, 10 mg or 20 mg/Kg/BW). Mice were sacrificed on 6 hours, 3 days and 14 days after the injection. Kidneys were fixed in the 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by refixation in the 1% osmium tetroxide solution. Dehydrated blocks were embedded in araldite mixture. The sections were cut on a LKB-V ultratome, and ultrathin sections stained with uranyl acetate and lead citrate were observed with JEM 100CX II electron microscope. The results were as follow: 1. Juxtaglomerular cell of the experimental groups showed some alterations, especially in the structures of protein synthesis including dilations and degradations of granular endoplasmic reticula, atrophy of Golgi complex, and numerous free ribosomes in the cytoplasm. 2. Juxtaglomerular cells treated groups showed a number of vacuoles, protogranules and some myelin figures in the cytoplasm, especially in the earlier groups. 3. Juxtaglomerular cells of treated groups, contained a large number of secretory granules showing variable electron densities and pleomorphism in later groups (2 weeks). From the above results, it was concluded that, the mercuric chloride or potassium bichromate induces acute renin release from juxtaglomerular cells of the mice, but many juxtaglomerular cells may secrete prematured secretory granules, or the synthetic system of the cell can not perform normal function.

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Enzymatic Synthesis of Meth.yl Fructoside by Immobilized Invertase (고정화 전화당 효소에 의한 메틸 프룩토시드의 합성)

  • 허주형;김해성
    • KSBB Journal
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    • v.8 no.4
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    • pp.313-319
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    • 1993
  • Methyl fructoside was synthesized from sucrose and methanol using an immobilized invertase. The enzyme was covalently bound by glutaraldehyde on porous silica coated with polyethyleneimine to give loading capacity of 120mg of invertase per one gram of dry porous silica and effective activity of 100U per one milligram of bound invertase. Polyethyleneimine coating imparted a hydrophillic character, good activity retention and high loading capacity to the surface of porous silica as well as hydrophillic microenviroment in the vicinity of bound invertase. The immobilized enzyme was formed into an alginate-enclosed silica bead to have enough activity for methyl fructoside synthesis from aqueous methanol-sucrose solution. Using the alginate-enclosed biocatalyst the yield of methyl fructoside was obtained as high as 55.9% from aqueous 30% (v/v) methanol and 0.291mo1/l sucrose with 2U/ml activity at $25^{\circ}C$, pH 4.8.

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Single-Protein Molecular Interactions on Polymer-Modified Glass Substrates for Nanoarray Chip Application Using Dual-Color TIRFM

  • Kim, Dae-Kwang;Lee, Hee-Gu;Jung, Hyung-Il;Kang, Seong-Ho
    • Bulletin of the Korean Chemical Society
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    • v.28 no.5
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    • pp.783-790
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    • 2007
  • The immobilization of proteins and their molecular interactions on various polymer-modified glass substrates [i.e. 3-aminopropyltriethoxysilane (APTS), 3-glycidoxypropyltrimethoxysilane (GPTS), poly (ethylene glycol) diacrylate (PEG-DA), chitosan (CHI), glutaraldehyde (GA), 3-(trichlorosilyl)propyl methacrylate (TPM), 3'-mercaptopropyltrimethoxysilane (MPTMS), glycidyl methacrylate (GMA) and poly-l-lysine (PL).] for potential applications in a nanoarray protein chip at the single-molecule level was evaluated using prismtype dual-color total internal reflection fluorescence microscopy (dual-color TIRFM). A dual-color TIRF microscope, which contained two individual laser beams and a single high-sensitivity camera, was used for the rapid and simultaneous dual-color detection of the interactions and colocalization of different proteins labeled with different fluorescent dyes such as Alexa Fluor® 488, Qdot® 525 and Alexa Fluor® 633. Most of the polymer-modified glass substrates showed good stability and a relative high signal-to-noise (S/N) ratio over a 40-day period after making the substrates. The GPTS/CHI/GA-modified glass substrate showed a 13.5-56.3% higher relative S/N ratio than the other substrates. 1% Top-Block in 10 mM phosphate buffered saline (pH 7.4) showed a 99.2% increase in the blocking effect of non-specific adsorption. These results show that dual-color TIRFM is a powerful methodology for detecting proteins at the single-molecule level with potential applications in nanoarray chips or nano-biosensors.

Pinopode Development 2-days after Oocyte Retrieval in the Human IVF Patients (체외수정 환자에서 난자회수 2일째의 자궁 내막의 Pinopode의 발달)

  • Lee, Kyung-Ah;Han, Sei-Yul;Choi, Dong-Hee;Lee, Woo-Sik;Yoon, Tae-Ki;Cha, Kwang-Yul
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.1
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    • pp.51-58
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    • 1998
  • 본 연구는 체외수정 프로그램에 참여하는 환자에 있어서 난자회수 이틀째의 자궁내막의 발달상태를 알아보기 위하여 pinopode의 발달상태, 에스트로젠 및 프로제스테론 수용체의 발현을 관찰하였다. 생검한 자궁내 막 조직 을 양분하여, 절반은 전사전자 현미경 (scanning electron microscope)으로 pinopode를 관찰하기 위하여 2.5% glutaraldehyde와 2% paraformaldehyde로 고정하였고, 나머지 절반은 dating 및 스테로이드 수용체의 면역조직화학적 측정 (immunocytochemistry)을 위하여 10% formalin으로 고정하였다. 모두 12명의 환자중 8명에서 pinopode가 관찰되었으며, pinopode 발달이 관찰되지 않은 환자들은 hCG 주사를 맞는 날의 estradiol (E2)의 혈중농도가 600 pg/mL이하로 낮았다. 본 연구의 결과로부터 자궁내막의 발달상태를 알아보기 위해서는 지금까지 일반적으로 사용되어 오던 dating이나 스테로이드 수용체의 면역조직화학적 측정법 이외에도 pinopode를 관찰함으로써 조금 더 정확한 진단을 할 수 있으리라고 사료되며, pinopode의 발달은 E2의 혈중농도와 관계가 있을 것으로 추정된다.

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Characteristics of Lactose Hydrolysis by Immobilized β-Galactosidase on Chitosan Bead (Chitosan 담체에 고정화된 β-galactosidase에 의한 유당 분해 특성)

  • Kang, Byung-Chul
    • Journal of Life Science
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    • v.21 no.1
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    • pp.127-133
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    • 2011
  • ${\beta}$-Galactosidase was immobilized on chitosan bead by covalent bonding using glutaraldehyde. The characteristics of the immobilized enzyme were investigated. Maximum immobilization yield of 75% was obtained on chitosan bead. Optimum pH and temperature for the immobilized enzyme was 7.0 and $50^{\circ}C$, respectively. The immobilized enzyme showed a broader range of pH and temperature compared to a free one. A mathematical model for the operation of the immobilized enzyme in a packed-bed reactor was established and solved numerically. Under different inlet lactose concentrations and feed flow rate conditions, lactose conversion was measured in a packed-bed reactor. The experimental results of continuous operation in a packed-bed reactor were compared to theoretic results using Michaelis-Menten kinetics with competitive product inhibition and external mass transfer resistance. The model predicted the experimental data with errors less than 5%. Process optimization of continuous operation in a packed-bed reactor was also conducted. In a recirculation packed-bed operation, conversion of lactose was 97% in 3 hours. In a continuous packed-bed operation, the effect of flow rate and initial lactose concentration was investigated. Increasing flow rates and initial lactose concentration decreased the conversion of substrate.

Regenerative Effects of Alloplastic Grafts in Rat Periodontal Fenestration Defects (백서 치조골 천공결손부에서의 합성골 이식재의 재생효과)

  • Gang, Yun-Kyung;Park, Joon_Bong;Kwon, Young-Hyuk;Herr, Yeek
    • Journal of Periodontal and Implant Science
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    • v.31 no.2
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    • pp.317-332
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    • 2001
  • The purpose of this study was to investigate effects of the natural coral(NC) and the hydroxyapatite/calcium sulfate hemihydrate(HA/CS) on an early stages of wound healing in the rat periodontal fenestration defects. In this experiment, twelve male rats(Mean : 520g in BW) aged 8 to 9 months were used. Experimental periodontal fenestration defects were surgically created with tapered fissure bur at the buccal surface of the left mandibular 1st, 2nd molars. The buccal aspects of molar roots were carefully denuded of their periodontal ligament through a bony window created in the left mandibles of rats under general anesthesia. Each experimental periodontal fenestration defect was grafted with natural coral and HA/CS, randomly. An area without bone graft was assigned for negative control group. At 10,35 days, rats were serially sacrificed via intracardiac perfusion with 2.5% glutaraldehyde and specimens were processed with Hematoxylin-Eosin stain for light microscopic evaluation. The results of this study were as follows : 1. The defect areas were filled with dense connective tissues at 10 days in control group. But in the test(NC, HA/CS)groups, the connective tissues around graft materials were formed more loosely and the response of inflammation by graft materials itself was not found. 2. The defect areas were filled with new osteoid tissues and new cementum was not formed on the cut root surface at 35 days in the control group. 3. New osteoid tissue formation was more prominent at 35 days in control than test groups. 4. The NC and HA/CS particles were encapsulated by loose connective tissues at 10 days and by dense connective tissues at 35 days, respectively. 5. In the test groups, resorption of graft particles was not found through the experimental time. From the above results, natural coral and hydroxyapatite/calcium sulfate hemihydrate may be biocompatible and osteoconductive and have a weak adverse reaction to the periodontal tissues.

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Drug Release Behavior and Degradability of Microspheres Prepared using Water-Soluble Chitosan (수용성 키토산으로 제조한 미세구의 분해성과 약물 방출 거동)

  • 장미경;최창용;김원석;정영일;나재운
    • Polymer(Korea)
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    • v.28 no.4
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    • pp.291-297
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    • 2004
  • Water-soluble chitosan micro spheres were prepared by emulsification of chitosan solution in mineral oil followed by cross linking reaction with different amount of the cross linking agent (glutraraldehyde), different chitosan concentration. Then, the physicochemical properties such as morphological change by degradation, drug loading efficiency, and drug release profiles were investigated with the drug loaded water-soluble chitosan microspheres. Norfloxacin loaded water-soluble chitosan micro spheres showed excellent drug entrapping capacities without burst release caused by surface bound drug. The absence of the surface bound drug also confirmed by X-ray diffraction study. Degradation and drug release studies showed that the amount of the crosslinking agent played a crucial role for drug loading, release and degradation. The water-soluble chitosan micro spheres showed more sustained drug release profiles with slower degradation and larger particle size by increasing crosslinking agent.