• Title/Summary/Keyword: Frozen sections

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A Freeze-fracture Study on the Odontoblast of Dental Pulp in the Rat Incisor (흰쥐 절치치수의 Odontoblast에 관한 Freeze-Fracture 연구)

  • Kim, Myung-Kook
    • Applied Microscopy
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    • v.16 no.2
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    • pp.1-13
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    • 1986
  • 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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Morphological Studies of the Central Neural Pathways to the Pancreas, Sanyinjiao(Sp6) and Yinlingquan(Sp9) using Pseudorabies Virus (Pseudorabies virus를 이용한 췌장과 삼음교(三陰交)(Sp6), 음릉천(陰陵泉)(Sp9)에서 투사되는 중추 신경로에 관한 형태학적 연구)

  • Kim, Cheol-Han;Lee, Su-Kyung;Yeom, Seung-Ryong;Kwon, Young-Dal;Song, Yung-Sun
    • Journal of Korean Medicine Rehabilitation
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    • v.19 no.1
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    • pp.23-38
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    • 2009
  • Objectives : The purpose of this morphological studies was to investigate the relations between Sanyinjiao(Sp6), Yinlingquan(Sp9) and pancreas of rats using peudorabies virus(PRV). Methods : We observed labeled neurons following the injection of PRV, Bartha strain, into the Sanyinjiao(Sp6), Yinlingquan(Sp9) and pancreas of rats. After survival times of 4 days following the injection of PRV, the rats were perfused, and their spinal ganglia, spinal cord and brain stem were frozen sectioned($35{\mu}m$). These sections were strained by PRV immunohistchemical staining methods and observed with light microscope. Results : The results were as follows. 1. In the spinal ganglia, the overlap areas of PRV labeled neurons projecting to Sanyinjiao(Sp6), Yinlingquan(Sp9) and pancreas were observed in T10-13 dorsal root ganglia. 2. In the spinal cord, the overlap areas of PRV labeled neurons projecting to Sanyinjiao(Sp6), Yinlingquan(Sp9) and pancreas were lamina I, IV, V, VII, IX, X, intermediolateral nucleus(IML), intermediomedial nucleus(IMM) in thoracic segments. In lumbar segments, the overlap areas of PRV labeled neuron were lamina I, IV, V, VI, IX, X and IMM. In sacral segments, the overlap areas of PRV labeled neuron were lamina I, IV, V, VI, VII, IX, X. 3. In the brain, the overlap areas of PRV labeled neurons projecting to Sanyinjiao(Sp6), Yinlingquan(Sp9) and pancreas were area postrema, nucleus tractus solitarius, caudoventrolateral reticular nu., medullary reticular nu., lateral paragigantocellular nu., C3 adrenalin cells, gigantocellular nu., raphe pallidus nu., raphe obscurus nu., ambiguus nu., raphe magnus nu., pontine reticular formation, A5 cell group, subcoeruleus nu., locus coeruleus, Barringnton's nu., $K{\ddot{o}}lliker$-Fuse nu., dorsal raphe nu., Edinger-Westphal nu., central gray matter, perifornical nu., dorsomedial hypothalamic nu., arcuate nu., lateral hypothalamic nu., paraventricular hypothalamic nu., hindlimb area. Conclusions : In conclusion, these results suggest that the interrelationship of meridian(spleen meridian), acupoints(Sp6 and Sp9) and viscera(pancreas) may be related the central autonomic centers.

A Case of Extensive Stage Small Cell Lung Cancer Presenting as an Acute Appendicitis with Perforation (전이성 병소에 의해 발생한 급성 충수염의 천공으로 진단된 전신병기 소세포폐암 1예)

  • Shin, Dong Won;Choi, Moon Han;Park, Seung Sik;Park, Sung Woo;Kim, Ki Up;Jang, An Soo;Park, Choon-Sik;Lim, Cheol Wan;Ko, Eun Suk;Paik, Sang Hyun;Kim, Do Jin
    • Tuberculosis and Respiratory Diseases
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    • v.65 no.3
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    • pp.230-234
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    • 2008
  • The incidence of appendiceal metastatic cancer is quite low. In particular, in small cell lung cancer, there is a very low incidence of a metastasis to the appendix. A 75-years old man with right lower quadrant pain, cough and sputum was transferred to our hospital. Abdominal CT revealed acute appendicitis with a perforation. The patient underwent surgery. The frozen sections of the tissue obtained during surgery, indicated a malignancy, but a right hemicolectomy was not performed due to the patient's poor general condition. The histology findings of the appendix were identified as a small cell carcinoma. The abdominal CT scan and chest x-ray at admission day showed a mass in the right lower lobe, and a further evaluation of the lesion was performed including positron emission tomography and flexible bronchoscopy with a biopsy. The pathology findings of the lung mass were also small cell lung cancer. The specimens from both sites stained positive for cytokeratin, cluster designation 56, synaptophysin, chromogranin-A and thyroid transcription factor 1. It was concluded that the appendiceal small cell cancer originated from the lung.

Enzyme-Histochemical Study of Philtral Orbicularis Oris Muscle Fiber Types in Korean Male Cadaver (한국인 성인남성 사체에서 시행한 인중 구륜근 섬유들의 효소-조직화학적 분석)

  • Yu, Myung-Sook;Park, Jung-Min;Lee, Hee-Su;Lee, Suk-Keun;Kang, Ji-Young;Eo, Mi-Young;Lee, Jong-Ho;Kim, Soung-Min
    • Korean Journal of Cleft Lip And Palate
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    • v.12 no.2
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    • pp.47-56
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    • 2009
  • The orbicularis oris muscle (OOM) is a very important muscle that originate from the second branchial arch and is innervated by the facial nerve. The aim of this study was to elucidate distribution types of two muscle fibers that composing OOM by using enzyme-histochemical examinations and tried to make a basis for a clinical application. The fresh frozen tissues from the superior and inferior portions of the OOM were taken from post mortem 65-year-old Korean male adult. Total five different sagittal sections were used on the midline of the philtrum, the middle portion of lower lip, the mouth corner, and each midlateral side of upper and lower mouth. We used enzyme-histochemical staining such as Periodic Acid-Schiff (PAS), Succinic Dehydrogenase (SDHase), reduced Nicotinamide Adenine Dinucleotide-Tetrazolium Reductase (NADH-TR), Adenosine Triphosphatase (ATPase) in pH 9.4, 4.6 and 4.3, and Modified Gomori Trichrome. There were about 30.24 % type 1 muscle fiber and 65.40 % type 2 muscle fiber in the midline of the philtrum (p < 0.05). Enzyme-histochemical staining is very useful and innovative method to elucidate characteristics of muscle fibers. We expect that chiloplasty and reconstruction of the lip portions for cleft lip patients, based on these results, are better to recovery function and aesthetic. However, we have some problems as an intramuscular variability and the inter-individual variation etc. Therefore we have to make progress these studies continuously to overcome these problems.

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Cytochemical Measurement of Lysosomal Responses in the Digestive Cells of Wild Pacific Oyster, Crassostrea gigas on the South Coast of Korea (세포화학적 방법을 이용한 남해안 조간대에 서식하는 참굴(Crassostrea gigas) 소화맹낭 세포 내 리소솜 활동 측정에 관한 연구)

  • Jeung, Hee-Do;Lee, Jee-Yeon;Hong, Hyun-Ki;Kang, Hyun-Sil;Kim, Young-Ok;Choi, Kwang-Sik
    • Ocean and Polar Research
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    • v.34 no.2
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    • pp.151-163
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    • 2012
  • Digestive cells of the bivalves have a highly developed lysosomal system and the system is known to be sensitive to changes in environmental qualities. In this study, we measured lysosomal responses of the digestive cells in wild oyster, Crassostrea gigas using frozen section. Oysters were collected in June 2010 from intertidal areas in the inner and outer bay of Gwangyang off the south coast of Korea. From the tissue sections, we measured the digestive cell lysosomal membrane stability (LMS), level of neutral lipids (NL), lipofuscin (LF) and the digestive gland atrophy (DGA). The DGA and condition index of oysters from the inner bay were significantly lower (P<0.05). The statistical test indicated that LMS levels of oysters in the inner bay and the outer bay were not significantly different since a shorter activity was displayed by oysters from the inner bay than that of oysters in outer bay. The LF deposition level of the oysters in the inner bay displayed significantly higher levels than the outer bay (P<0.05). In contrast, the NL accumulation measured from oysters in outer bay was significantly higher than the level observed in the inner bay (P<0.05). Different levels of DGA and LF that were observed in the inner and outer bays were thought to be associated to different level of environmental contamination and these two assays are considered to be good biomarkers.

Central Neural Pathways Projecting to the Acupoints B62 and K6 Using the Neural Tracer (신경추적자를 이용한 신맥과 조해에서 투사되는 신경원의 표지부위에 대한 연구)

  • Kim, Su-Hyun;Lee, Chang-Hyun;Yuk, Sang-Won;Lee, Kwang-Gyu;Lee, Tae-Young;Lee, Sang-Ryoung
    • Journal of Acupuncture Research
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    • v.18 no.2
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    • pp.51-66
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    • 2001
  • The purpose of this morphological studies was to investigate the central neural pathway projecting to the acupoints $B_{62}$ and $K_6$ using the neuroanatomical method following injection of transsynaptic neurotropic virus, pseudorabies virus(PRV-Ba and PRV-Ga) into the $B_{62}$ and $K_6$. After survival times of 96 hours following injection into the twenty rats with PRV-Ba(Bartha strain) and PRV-Ga(Bartha strain, ${\beta}$-galacidodase insertion). They were perfused, and their spinal cord and brain were frozen sectioned($30{\mu}m$). These sections were stained by X-gal histochemical and PRV immunohistochemical staining method, and observed with light microscope. The results were as follows : 1. In spinal cord, overlaped PRV-Ba and PRV-Ga labeled neurons projecting to the $B_{62}$ and $K_6$ were founded in thoracic, lumbar and sacral spinal segments. In thoracic spinal segments, Densely labeled areas were founded in lamina IV, V, VII(intermediolateral nucleus) and X areas. In lumbar segemnts, labeled areas were founded in lamina II, IV, V and X areas. In sacral spinal segments, labeled areas were founded in lamina IV, V and VI areas. 2. In brain, overlaped PRV-Ba and PRV-Ga labeled neurons projecting to the $B_{62}$ and $K_6$ were founded in the $A_1$ noradrenalin cells/$C_1$ adrenalin cells/caudoventrolateral reticular nucleus, rostroventrolateral reticular nuclens, nucleus tractus solitarius, area postrema, raphe obscurus nucleus, raphe paltidus nucleus, raphe magnus nucleus, lateral paragigantoceltular nucleus, lateral rcticular nucleus, gigantocellular nucleus, locus coeruleus, subcoeruleus nucleus, motor trigeminal nucleus, Kolliker-Fuse nucleus, $A_5$ cell group, central gray matter, oculomotor nerve, paraventricular hypothalamic nucleus, median eminence, amygdaloid nucleus, frontal cortex, forelimb area, hindlimb area, 1, 2 areas of parietal cortex and granular and agranular cortex. This results were suggest that overlaped PRV-Ba and PRV-Ga labeled areas projecting to the $B_{62}$ and $K_6$ may be related to the emotional relay pathway in the central autonomic center.

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Neuroanatomical Studies on the Acupoints Related to the Large Intestine (대장(大腸)과 관련(關聯)된 경혈(經穴)들의 신경해부학적(神經解剖學的) 연구(硏究))

  • Kang, Chang-Soo;Lee, sang-ryoung;Lee, Chang-Hyun;Nam, Yong-Jae;Lee, Kwang-Gyu
    • Journal of Acupuncture Research
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    • v.17 no.2
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    • pp.95-117
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    • 2000
  • The purpose of this morphological studies was to investigate the relation between the meridian, acupoints and viscera using neuroanatomical tracers. The common locations of the spinal ganglia, sympathetic chain ganglia, spinal cord and brain projecting to the large intestine meridian were observed following injection of transganglionic tracer, WGA-HRP and transsynaptic neurotropic virus, pseudorabies virus(PRV), Bartha strain(Ba) and PRV-Ba-Gal (Galactosidase)) into the the large intestine(cecum, colon and rectum), ST37 and LI4. After survival times of 96 hours following injection into the thirty rats with WGA-HRP, PRV-Ba and PRV-Ba-Gal. They were perfused, and their spinal ganglia, sympathetic chain ganglia, spinal cord and brain were frozen sectioned($30{\mu}m$). These sections were stained by HRP and X-gal histochemical and PRV immunohistochemical staining method, and observed with a light microscope. The results were as follows : 1. WGA-HRP labeled neurons innervating the large intestine were observed bilaterally within the T13-L4 sympathetic chain ganglia, and T9-11 spinal ganglia. WGA-HRP labeled neurons innervating ST37 were observed within the L3-5 sympathetic chain ganglia, and L2-4 spinal ganglia. WGA-HRP labeled neurons innervating LI4 were observed in the middle cervical ganglion and stellate ganglion, and C5-8 spinal ganglia. 2. In spinal cord, PRV-Ba labeled neurons projecting to the large intestine, ST37 and LI4 were found in thoracic, lumbar and sacral spinal segments. Densely labeled areas of each spinal cord segment were founded in lamina N, V, VII(intermediolateral nucleus), Ⅸ, X and dorsal nucleus. 3. In medulla oblongata, PRV-Ba and PRV-Ba-Gal labeled neurons projecting to the large intestine, ST37 and LI4 were commonly found in the A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nucleus, dorsal motor nucleus of vagus nerve, nucleus tractus solitarius, raphe obscurus nucleus, raphe pallidus nucleus, raphe magnus nucleus and gigantocellular nucleus. 4. In pons, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in locus coeruleus, Kolliker-Fuse nucieus and A5 cell group. 5. In midbrain, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in central gray matter. 6. In diencephalon, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in paraventricular hypothalamic nucleus. These results suggest that PRV-Ba and PRV-Ba-Gal labeled common areas projecting to the large intestine may be correlated to that of the large intestine meridian, ST37 and LI4. Especially, These morphological results provide that interrelationship of meridian-acupoints -viscera may be related to the central autonomic pathways.

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Localization of the Neurons Projecting to the Gallbladder Meridian (족소양담경(足少陽膽經)에서 투사(投射)되는 신경원(神經元)의 표지부위(標識部位)에 대한 연구(硏究))

  • Ryuk Sang-Won;Lee Kwang-Gyu;Lee Sang-Ryoung;Kim Jum-Young;Lee Chang-Hyun;Lee Bong-Hee
    • Korean Journal of Acupuncture
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    • v.17 no.1
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    • pp.101-121
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    • 2000
  • The purpose of this morphological studies was to investigate the relation to the meridian, acupoint and nerve. The common locations of the spinal cord and brain projecting to the the gallbladder, GB34 and common peroneal nerve were observed following injection of transsynaptic neurotropic virus, pseudorabies virus(PRV), into the gallbladder, GB34 and common peroneal nerve of the rabbit. After survival times of 96 hours following injection of PRV, the thirty rabbits were perfused, and their spinal cord and brain were frozen sectioned($30{\mu}m$). These sections were stained by PRV immunohistochemical staining method, and observed with light microscope. The results were as follows: 1. In spinal cord, PRV labeled neurons projecting to the gallbladder, GB34 and common peroneal nerve were founded in thoracic, lumbar and sacral spinal segments. Densely labeled areas of each spinal cord segment were founded in lamina V, VII, X, intermediolateral nucleus and dorsal nucleus. 2. In medulla oblongata, The PRV labeled neurons projecting to the gallbladder, GB34 and common peroneal nerve were founded in the A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nucleus, rostroventrolateral reticular nucleus, medullary reticular nucleus, dorsal motor nucleus of vagus nerve, nucleus tractus solitarius, raphe obscurus nucleus, raphe pallidus nucleus, raphe magnus nucleus, gigantocellular nucleus, lateral paragigantocellular nucleus, principal sensory trigeminal nucleus and spinal trigeminal nucleus. 3. In Pons, PRV labeled neurons were parabrachial nucleus, Kolliker-Fuse nucleus and cochlear nucleus. 4. In midbrain, PRV labeled neurons were founded in central gray matter and substantia nigra. 5. In diencephalon, PRV labeled neurons were founded in lateral hypothalamic nucleus, suprachiasmatic nucleus and paraventricular hypothalamic nucleus. 6. In cerebral cortex, PRV labeled neuron were founded in hind limb area.This results suggest that PRV labeled common areas of the spinal cord projecting to the gallbladder, GB34 and common peroneal nerve may be first-order neurons related to the somatic sensory, viscero-somatic sensory and symapathetic preganglionic neurons, and PRV labeled common area of the brain may be first, second and third-order neurons response to the movement of smooth muscle in gallbladder and blood vessels.These PRV labeled neurons may be central autonomic center related to the integration and modulation of reflex control linked to the sensory system monitoring the internal environment, including both visceral sensation and various chemical and physical qualities of the bloodstream. The present morphological results provide that gallbladder meridian and acupoint may be related to the central autonomic pathways.

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Analyzing Driving Behavior, Road Sign Attentiveness and Recognition with Eye Tracking Data (운전자 시각행태 및 주행행태 분석기반의 결빙주의표지 개발연구)

  • Lee, Ghang Shin;Lee, Dong Min;Hwang, Soon Cheon;Kwon, Wan Taeg
    • The Journal of The Korea Institute of Intelligent Transport Systems
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    • v.20 no.6
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    • pp.117-132
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    • 2021
  • Due to the terrain in Korea, there are many road sections passing through mountainous areas. During the winter, there is a higher risk of traffic accidents, due to black ice caused by the lack of sunlight. Despite domestic road freezing safety measures, accidents caused by road freezing results in severe traffic accidents. Under these considerations, this study analyzed whether traffic safety signs that change in response to the external temperature help drivers recognize frozen road segments. The study was conducted through analysis of the effect of the signs on a driver's perspective. For the signs under development, out of the signs designed by experts, the sign design which received the highest visibility and effectiveness evaluation ratings from the general public was selected. The sign was implemented through Virtual Reality (VR) and installed on the right side of the road to analyze the effect on gazing and driving behavior. As a result of analyzing the driver's driving behavior, a speed reduction of about 7km/h or more was found in the sign section. Therefore, It was found that the existence of the sign had a strong relationship with the rate of the drivers' speed reduction.

The Distribution of ${\gamma}{\delta}$ T Cells in Tuberculous Lymphadenopathy (결핵성 림프절에서 ${\gamma}{\delta}$ T 림프구의 분포에 관한 연구)

  • Shim, Tae-Sun;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.5
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    • pp.484-488
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    • 1994
  • Background : The antigen-specific receptor on the surface of most peripheral T lymphocytes is a disulfide-linked heterodimer composed of $\alpha$ and $\gamma$ subunits, noncovalently associated with CD3 polypeptides. Recently, a novel type of CD3-associated heterodimer was described on a T cell subset that does not express CD4 or CD8 molecules. This second type of TCR dimer is composed of chains encoded for by the $\gamma$- and $\delta$-TCR genes. These cells may exert both cytotoxic and lymphokine producing functions. Although it was reported that some ${\gamma}{\delta}$-TCR might recognize an MHC-linked determinant, the funεtion or physiologic ligand for this new receptor is not yet clear. It was found that ${\gamma}{\delta}$-TCR can react with 65 kD heat shock protein of M. tuberculosis, which suggests the possible protective role of ${\gamma}{\delta}$ T lymphocytes against tuberculosis. In our previous study, there was neither the increase in number nor the functional activation of ${\gamma}{\delta}$ T cells in the peripheral blood from patients with pulmonary tuberculosis. Now we report the distribution of ${\gamma}{\delta}$ T cells in the regional sites of M. tuberculosis infection, especial1y tuberculous lymphadenitis. Methods : Lymph nodes from patients with pathologically-proven tuberculous lymphadenopathy (n=5) and reactive hyperplasia (n=3) were used. Tissues were frozen in liquid nitrogen immediately after removal and stored below $-70^{\circ}C$. The cryostat sections of these frozen specimens were stained with anti-Leu-4 Ab, Identi-T TCR ${\delta}1$, and Identi-T ${\beta}F1$. The number of positively stained cells were counted at high power field. Results : The infiltration of ${\gamma}{\delta}$ T cells was significantly higher in the lymph nodes from patients with tuberculous lymphadenopathy than that with reactive hyperplasia ($16.3{\pm}10.3%$ vs. $1.7{\pm}1.5%$). Conclusion : These results suggest that ${\gamma}{\delta}$) T cells may play a role in the defense against M. tuberculosis infection, especially in the regional sites of infection.

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