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RISS 인기검색어
- 검색키워드
- 저자 : 김명국(Myung Kook Kim) (검색결과 83 건)
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EM Radioautographic Techniques에 관(關)한 연구(硏究) - Cork 방법(方法) -
김명국,Kim, Myung-Kook,Hassler, R. 한국현미경학회 1980 Applied microscopy Vol.10 No.1
Electron microscope radioautography introduced by Liquier-Milward (1956) is now used routinely in many laboratories. Most of the technical difficulties in specimen preparation have been overcome. This method is modified from loop method for improvement of EM radioautographic techniques. The advantages of this method are: 1. the use of single specimens on small corks and of a large wire loop, allows the experimenter to avoid the blemishes in the membrane; 2. the surfactant dioctyl sodium sulphosuccinate is added to diluted ILford L4, thus greatly prolonging the period of time over which good emulsion layers can be made; 3. corks can be handled in perspex holder which allows about 20 specimens to be developed simultaneously. The steps of the method comprise: 1. Cut ribbons of ultrathin sections of silver interference colour 2. Pick them up on formvar-coated 200 mesh grids 3. Prestaining of tissues 4. Coat the specimens with a thin layer of carbon by evaporation (30-60A) 5. Mount the specimens on corks (about 1cm apical diameter) using double-sided scotch tape 6. Emulsion coating; a. Take a 250m1 beaker, place it on the pan of a sliding weight balance and weigh it. Add 10 grams extra to the beam. Add pieces of ILford L4 emulsion to the beaker until the balance is swinging freely. Add the 20ml of distilled water that was previously measured out. b. Surfactant dioctyl sodium sulphosuccinate is added to diluted ILford L4. 7. Prepare a series of membranes of gelled emulsion with the wire loop and apply one to each cork-borne specimen. 8. Put the specimens away to expose by pushing the corks into short length of PVC tubing, each tube having a small hole in the side 9. Place the tubes in small boxes together with silica gel. 10. Exposure 11. Developer - Kodak Microdol X for 3 minutes 12. Fixer - A perspex holder can be manufactured which allows 20 specimens to be developed simultaneously. 12. Fixer - 30% sodium thiosulfate for 10 minutes 13. Examination with Siemens Elmiskop 1A electron microscope
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연령증가에 따른 흰쥐 삼차신경척수핵 중간부분에서의 신경연접구조의 변화에 관한 연구
김명국,김철위,백기석,임범순,Kim, Myung-Kook,Kim, Cheol-We,Paik, Ki-Suk,Lim, Bum-Soon 한국현미경학회 1998 Applied microscopy Vol.28 No.3
This study was performed to observe the morphological changes of the synaptic structures in the interpolar part of the spinal trigeminal nucleus of rat during aging. Transmission electron microscopy has been used to determine the r)umber of synapses, length of postsynaptic densities, number and area of axon terminals. Sprague-Dawley rat 3, 12, 24 and 36 months of age were used in this study. 1. The number of synapses was 51.7, 43.1, 28.4 and 16.8 in the 3, 12, 24 and 36 months of age respectively. Therefore, the number of synapses decreased gradually with age, but decreased significantly in the 24 and 36 months. 2. The length of postsynaptic densities was $30.2{\mu}m,\;23.6{\mu}m,\;10.4{\mu}m\;and\;4.9{\mu}m$ in the 3, 12, 24 and 36 months of age respectively. Therefore, the length of postsynaptic densities decreased gradually with age, but decreased significantly in the 24 and 36months. 3. The number of axon terminals was 84.3, 73.7, 51.4 and 26.6 in the 3, 12, 24 and 36 months of age respectively. Therefore, the number of axon terminals decreased gradually with age, but decreased significantly in the 24 and 36months. 4. The area of axon terminals was $76.1{\mu}m^2,\;64.1{\mu}m^2,\;29.9{\mu}m^2\;and\;13.8{\mu}m^2$ in the 3, 12, 24 and 36 months of age respectively. Therefore, the area of axon terminals decreased gradilally with age, but decreased significantly in the 24 and 36 months. The results suggest that there are the changes of the synaptic structures in the interpolar part of spinal trigeminal nucleus of rat during aging. These changes nay be concerned to the decreased function of mediating pain and temperature sensation in the face and oral cavity during aging.
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생쥐 상악치조부에서의 파골세포의 당단백 합성 및 이동에 관한 전자현미경 자기방사법적 연구
김명국,Kim, Myung-Kook 한국현미경학회 1992 Applied microscopy Vol.22 No.2
The pathway and time course of fucose-containing glycoprotein synthesis and intracellular translocation in osteoclasts of the mice maxillary alveolar bone were investigated by electron microscopic radioautography. Male Balb-C mice weighing 17gm were anesthetized with Nembutal and injected via the external jugular vein with 2.5 mCi of $L-[6-^{3}H]-fucose$ (specific activity 16.8 mCi/mmol) in 0.1 ml of sterile saline solution. At 5, 10, 20, 35 minutes and 8 hours after administration of the $^{3}H-fucose$, animals were killed by intracardiac perfusion of 30ml of 2% glutaraldehyde in a modified Tyroid solution, pH 7.4. The maxillae were then removed and further fixed in Karnovsky fixative for an additional 3-4 hours. After rinsing in 0.1M cacodylate buffer for 10 minutes, the maxillae were demineralized for 2 weeks at $4^{\circ}C$ in ethylene diamine tetra acetate containing 2% glutaraldehyde. The first interdental areas were mesiodistally sectioned into slices of 1mm thickness and postfixed in osmium tetroxide. Tissues were then dehydrated and embedded in Poly Bed. To prepare electron microscopic radioautography, the dipping method of Kopriwa (1973) was employed. Thin sections were coated with a crystalline monolayer of ILford $L_4$ photographic emulsion. After exposure for 4 months at $4^{\circ}C$, the sections were developed Kodak Microdol-X and Phenidon (for compact grains), fixed in 30% sodium thiosulfate, stained with uranyl acetate and lead citrate and examined in the electron microscope (JEOL 1200 EX). At 5, 10 and 20 minutes after injection, $^{3}H-fucose$ was concentrated in Golgi cisternae of the osteoblasts. By 35 minutes the labels were observed over small vesicles in the suprannclear area of osteoclasts. At 8 hours, numerous silver grains were located on the ruffled border and cell membrane of osteoclasts. These results indicate that fucose molecules are added in the Golgi apparatus and small vesicles appear to be responsible for translocation of the glycoproteins to the marginal portion of osteoblasts. The glycoproteins are distributed on the osteoclast cell surface and especially over the ruffled border.
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노화된 흰쥐 뇌 삼차신경주감각핵에 관한 전자현미경적 연구
김명국,Kim, Myung-Kook 한국현미경학회 1995 Applied microscopy Vol.25 No.1
The purpose of this study was to investigate the main sensory trigeminal nucleus in the aging rat brain by means of electron microscope. Male Sprague-Dawley rats, two (control group) and thirty six (aging group) months of age, were used. These animals were sacrificed by perfusion fixation with 2.5% glutaraldehyde-2.0% paraformaldehyde (0.1M phosphate buffer, pH 7.4) under sodium pentobarbital. The objective area was punched out with a sharp-edged metal cylinder of 0.8 mm in diameter. These blocks of tissue were then washed in 0.1M phosphate buffer, postfixed in 2% osmium tetroxide, dehydrated in a graded series of ethyl alcohol, and embedded in Epon 812. Thin sections were cut with Super Nova ultramicrotome, pick up on grids and double stained with lead citrate and uranyl acetate, and observed in JEOL 100B electron microscope. The results were as follows: 1. In the control group, the neuronal cell body of the main sensory trigeminal nucleus was filled with nucleus, Golgi complex, Nissl substance, mitochondria, microfilaments and microtubules. However, few Nissl substances are seen in neuronal cell body. Axoaxonic synapse, axodendritic synapse, axosomatic synapse, axospinous synapse, myelinated and unmyelinated nerve fibers were well organized around cell bodies. Neurons with abnormal changes were not seen. 2. In the aging group, the neuronal cell body of the main sensory trigeminal nucleus contained large number of lipofuscin granules, dense body and swollen mitochondria. Terminal boutons contained glycogen, crystal-like vesicle and membranous indicating first signs of degeneration. The dendrites were found to be in synaptic contact with altered axon terminals. Frequently axons filled with dark axoplasn and splitted myelin sheath were noticed.
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흰쥐 절치치수의 Odontoblast에 관한 Freeze-Fracture 연구
김명국,Kim, Myung-Kook 한국현미경학회 1986 Applied microscopy Vol.16 No.2
The purpose of this study was to investigate the morphology and intercellular junctions of the odontoblast of dental pulp in the rat incisor by means of the freeze fracture electron microscopy. Twenty male Sprague-Dawley rats weighing $150{\sim}200g$ were used. After being anesthetized by an intraperitoneal injection of 0.5 ml sodium pentobarbital per kg in body weight(60 mg/ml) the animals were perfused with 2.5% glutaraldehyde-2% paraformaldehyde fixative in 0.1 M cacodylate buffer, pH 7.2 through the ascending aorta for one hour. The incisors were carefully extracted from the jaws and demineralized by suspending them in 0.1 M EDTA in 3% glutaraldehyde (pH 7.2) for two weeks. After demineralization, the specimens were obtained from the portion divided into five equal parts. For freeze-fracture replication, demineralized tissues were infiltrated for several hours with 10%, 25% glycerol in 0.1M cacodylate buffer as a cryoprotectant and then frozen in liquid Freon 22 and stored in liquid nitrogen. Fracturing and replication were done in Balzers BAF 400D high-vacuum freeze-fracture apparatus at $-120^{\circ}C$ under routine $5X10^{-7}$ Torr vacuum. The tissue was immediately replicated with platinum unidirectionally at $45^{\circ}$ angle and reinforced with carbon at $90^{\circ}$ angle unidirectionally or by using a rotary stage. The replication process was monitored by a quartz-crystal device. The replicas were immersed in 100% methanol overnight. The tissue was then digested from the replica by clorox (laundry bleach), placed into 5% EDTA, and washed repeatedly with distilled water. The replicas were picked up on 0.3% formvar-coated 75 mesh grids and examined in the JEOL 100B electron microscope. The results were as follows; 1. Both in thin sections and freeze-fracture replicas, three types of intercellular junctions were recognizable in the plasma membrane of odontoblast: gap junction, tight junction and desmosome-like junction. 2. The nuclear pores were evenly distributed over the nuclear envelope. The pore complex formed a ring about 70 nm in diameter. 3. Gap junctions were found between odontoblasts as well as odontoblasts and neighbouring pulp cells (fibroblast, subodontoblastic cell process, nerve-like fibre). Gap junctions, which were round, ellipsoid and pear-shaped and 600 nm in diameter, were observed in the odontoblast. 4. Numerous round and ellipsoid gap junctions could be frequently seen on the plasma membranes in cell body and apical part of the odontoblasts. On the P face, the junctions were recognized as a cluster of closely packed particles, measuring about 9 nm in diameter, and on the E face, the junctions were recognized as a shallow grooves.
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