• Title/Summary/Keyword: Glutaraldehyde

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An Electron Microscopy of the Retina in the Gallus domesticus B. (家鷄 網膜의 電子顯微鏡的 硏究)

  • Paik, Kyung Ki;Choi, Choon Keun
    • The Korean Journal of Zoology
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    • v.15 no.2
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    • pp.71-85
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    • 1972
  • This investigation was undertaken to establish the ultrastructural organization of the retina in domestic fowl (Gallus domesticus B.) comparing with the ultrastructure that has been indicated in other Aves by several workers. The electron microscope observations were made on selected segments of retinal tissue prefixed for 2 hrs in 1.25% glutaraldehyde buffered with 0.2 M cacodylate at pH 7.2 and then postfixed in cold 1% osmium tetroxide in 0.4 M cacodylate buffer for 2 hrs. After postfixation, tissues were dehydrated in alcohol series, embedded in Epon 812 mixture from propylene oxide and stained with saturated uranyl acetate and $Pb(NO_3)_2$ solution. Specimens were examined with a Hitachi HS-7S electron microscope. The pigment epithelia cells contain numerous mitochondria with prominent dense granules and several changeful spaped Golgi bodies. The internal fine structure of the receptor outer segments revealed the characteristic stacks or arrays of bimembranous disks. The ellipsoid outer portion of the cone inner segments is composed of a tightly packed mass of extraordinarily large mitochondria. The outer limiting membrane is seen to contain many junctional complexes, the fibrillar material of which is electron-dense.

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Surface ultrastructures of the third-stage larvae of Gnathostoma hispidum (돼지악구충(Gnuhostoma hispidum) 제3기 유충의 표면 미세구조)

  • 손운목;이진하
    • Parasites, Hosts and Diseases
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    • v.34 no.3
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    • pp.169-176
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    • 1996
  • A scanning electron microscopic study was performed to observe the surface ultrastructures of the third-stage larvae of Gnathostoma hispidun. The early third-stage larvae (EL3) were collected from the viscera of Chinese loaches by the artificial digestion method . The advanced third-stage larvae (AdL3) were recovered from mice experimentally infected with EL3. Both larval worms were fixed with 2.5% glutaraldehyde, dehydrated in graded alcohol. dryad in critical point dryer, and coated with gold. The specimens were observed with a SEM (DS- l30C). On the head bulb of both larval stage, the mouth had a pair of lateral lips of equal size and of half moon shape. Each lip had a couple of labial papillae and a small amphid located between the two papillae. The hooklets on the head bulb had single-pointed tips and curved posteriorly. The cuticular spines of EL3 were larger and more densely distributed in the anterior area (about 1.8 Mm in length) and gradually decreased in size and number posteriorly. The cuticular spines in the anterior area of AdL3 were sharp-pointed and about 4.5 Mm in length, and those in the middle area were about 1.75 Mm. The velvety cuticular folds and dot-like cuticular spines were distributed in the posterior area. A cervical papilla was located between the 7th and 8th transverse striations. A dome-like body papilla was located at the posterior 1/4 of body. An ellipsoidal excretory pore was located between the 17th and 18th striations. From the above results, it is suggested that the characteristic SEM findings obtained from this study may be helpful on the species identification of larval Gncthostomn.

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Do Paneth Cells Regulate the Zinc Body Burden? (Zinc 대사와 관련된 Paneth 세포활성의 변화에 관한 조직화학적 연구)

  • Jo, Seung-Mook;Kim, Sung-Jun;Park, Seung-Kook;Kang, Tae-Cheon;Won, Moo-Ho
    • Applied Microscopy
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    • v.30 no.4
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    • pp.357-365
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    • 2000
  • Paneth cells have been suggested to contribute to the elimination of excess metals into the intestinal lumen. The purpose of this study wat to investigate the changes of the zinc pools in rats subjected to functional loading with zinc salt by mean of both light and electron microscopical autometallography (AMG). Wistar rats 4 were administrated with zinc chloride (20 mg/kg body weight) intraperitoneally dissolved in 1 ml distilled water. The control group received 1 ml saline IP. After further one hour the animals were transcardially perfused with 0.4% sodium sulphide dissolved in 0.1 M PB fellowed by 3% glutaraldehyde solution for 10 minutes. Pieces of ileum were frozen with solid $CO_2$ and sectioned on a cryostat. The sections $(20{\mu}m)$ were autometallographically developed. Sections selected for EM were reembedded on top of a blank Epon block, from which ultrathin sections (100 nm) were cut. The ultrathin sections were double stained with uranyl acetate (30 min) and lead citrate (5 min), then examined under electron microscope. Studies of comparable sections from control and zinc loaded animals with the AMG selenium method gave quite different results. The control animals demonstrated a weakly positive staining in the cytoplasm of the Paneth cells. In the electron microscope the AMG silver grains were found to be located in the cytoplasm, while the electron dense secretary granules and other cell organelles were void of staining. Few AMG grains were located at the apical surface of the Paneth cells. In sections from zinc loaded rats, the AMG grains were seen in abundance in the lumen of the Lieberkuhn crypts at light microscopic levels. At EM levels the zinc revealing silver grains were located in the cytoplasm as in the controls, but much more AMG grains were shifted into the secretary granules. Furthermore, profound AMG grains were found in the lumen of the crypts and surrounding vessels. And a few grains were seen in the endothelium. The AMG technique demonstrated a pattern of AMG grains in the Paneth cells that strongly suggests a transport of zinc ions through these cells.

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Tegumental ultrastructure of juvenile and adult Echinostoma cinetorchis (이전고환극구흡충 유약충 및 성충의 표피 미세구조)

  • 이순형;전호승
    • Parasites, Hosts and Diseases
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    • v.30 no.2
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    • pp.65-74
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    • 1992
  • The tegumental ultrastructure of juvenile and adult Echinostoma cinetorchis (Trematoda: Echinostomatidae) was observed by scanning electron microscopy. Three-day (juvenile) and 16-day (adult) worms were harvested from rats (Sprague-Dawley) experimentally fed the metacercariae from the laboratory-infected fresh water snail, Hippeutis cantori. The worms were fifed with 2.5% glutaraldehyde, processed routinely, and observed by an ISI Korea DS-130 scanning electron microscope. The 3-day old juvenile worms were elongated and ventrally curved, with their ventral sucker near the anterior two-fifths of the body. The head crown was bearing 37∼38 collar spines arranged in a zigzag pattern. The lips of the oral and ventral suckers had 8 and 5 type II sensory papillae respectively, and bewteen the spines, a few type III papillae were observed. Tongue or spade-shape spines were distributed anteriorly to the ventral sucker, whereas peg-like spines were distributed posteriorly and became sparse toward the posterior body. The spines of the dorsal surface were similar to those of the ventral surface. The 16-day old adults were leaf-like, and their oral and ventral suckers were located very closely. Aspinous head crown, oral and ventral suckers had type II and type III sensory papillae, and numerous type I papillae were distributed on the tegument anterior to the ventral sucker. Scale-like spines, with broad base and round tip, were distributed densely on the tegument anterior to the ventral sucker but they became sparse posteriorly. At the dorsal surface, spines were observed at times only at the anterior body. The results showed that the tegument of E. cinetorchis is similar to that of other echinostomes, but differs in the number and arrangement of collar spines, shape and distribution of tegumenal spines, and type and distribution of sensory papillae.

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The Ultrastructure of Leydig Cells in the Testis of Korean Native Chickens (한국 재래 닭 고환 간질세포의 미세구조에 관한 연구)

  • Jang, B.G.;Tae, H.J.;Choi, C.H.;Park, Y.J.;Yang, H.H.;Kim, N.S.;Park, S.Y.;Kang, H.S.;Park, B.Y.;Lee, Y.H.;Ahn, D.C.;Kim, I.S.
    • Korean Journal of Poultry Science
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    • v.33 no.3
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    • pp.181-188
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    • 2006
  • Changes in the fine structure of testicular Leydig cell from hatching to adulthood were studied in Korean native chickens of 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 24, 28, 32, 44, 52 and 64 weeks (n=13 chickens per group) of age. The objective of this study were to elucidate Leydig cell ultrastructure during testicular development. Testes of chickens were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using $1{\mu}m$ sections stained with methylene blue-azure II, qualitative and quantitative(stereological) morphological studies were performed. The ultrastructural changes of the Leydig cell were investigated by ultrathin section with the transmission electron microscope. The stages of the Leydig cell development described focus on mitochondria, endoplasmic reticulum, and lipid droplets which are involved in androgens as fullows. 1) Approaching puberty. The closely packed Leydig cells and sparse intercellular space. The nucleus occupied a large portion of the Leydig cell volume. The population of Leydig cells contained two types of cells that differed in the appearance of their nuclei which were either highly electron-opaque or relatively electron-lucid. The cytoplasm was characterized by large amounts of lipid droplets, relatively few spherical mitochondria, and sparse smooth endoplasmic reticulum. 2) Puberty to adult. The Leydig cells which display features compatible with significant androgen synthesis: large volume of cytoplasm containing extended smooth endoplasmic reticulum, abundant mitochondria, and reduction of lipid droplets.

Tissue Failure of the Low-Profile lonescu-Shiley Pericardial Valve in Mitral Position (승모판에서의 단고형 이오네스큐판막의 조직실패)

  • 김종환
    • Journal of Chest Surgery
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    • v.30 no.7
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    • pp.670-676
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    • 1997
  • The structural failure of the glutaraldehyde-treated xenograft valves has been the primary concern about the limited durability as predicted from the begimling of clinical use, and long-term follow-up has shown a significant incidence of primary tissue failure(PTF) from both biological and mechanical reasons. Twenty-seven patients with the low-profile lonescu-Shiley valves explanted from mitral position for PTF(Group III) were studied on the patient characteristics and valve pathology, and the results were compared with the matched observations of the Haycock(Group I) and of the standard-profile lonesiu-Shiley valves(Group II). Patients were aged 16 to 56 years(mean, 38.0$\pm$ 11.0 years), and the size of the failed mitral bioprosthesis was 30.8$\pm$ 1.3 mm. The hemodynamic consequences were stenosis in 29.6%, insufficiency in 44.4%, mixed steno-insufficiency in 14.8%, together with normal function for the rest of patients of prophylactic re-replacement. Pathology revealed calcification with or without tissue damage in 63.0% and tissue damage with or without calcification in 58.l%, in contrast with the observations of predominant tissue damage(76.8%) over calcification in Group I and of calcification(76.1%) over tissue damage in group II. Although dystrophic calcification has long and repeatedly dealt with patient's young age as a determinant of valve durability, such a characteristic evidence was not reached even in patients with calcified valves. Moreover, the prolonged explantation p riods from the studied on the previous report suggested strongly yet possibly evolving destructive processes among the valves in the remaining patients, and awaits further follow-up. In conclusion, PTF of the xenograft valves seems to result from more complicated biologic and metabolic reasons as well as more complex mecharical factors than the reported, and newer generation prostheses, with tissue preservation with glutaraldehyde, do not likely to provide decisive improvement in the occurrence of structural failurebioprostheses is generally limited to the highly aged.

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Aortic Valvuloplasty Using Leaflet Extension Technique (판막첨 연장술을 이용한 대동맥판막 성형술의 중기성적 평가)

  • Ahn, Hyuk;Kim, Hyun-jo
    • Journal of Chest Surgery
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    • v.30 no.7
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    • pp.656-662
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    • 1997
  • Aortic valvuloplasty has recently gained attention as an attractive alternative procedure for aortic valvular disease. Between March 1995 to August 1996, 14 patients with pure aortic regurgitation(AR) underwent aortic alvuloplasty using leaflet extension with glutaraldehydepreserved autologous pericardium. There were 11 males and 3 females, and the mean age was 34.8 $\pm$ 15.3 years. Preoperative echocardiography and cardiac catheterization revealed that the degree of AR was mean 3.4$\pm$0.65, and more than moderate degree of mitral regurgitation(MR) were detected in 4 patients. In 12 patients, 3 leaflets were extended and in another 2 patients only one deformed leaflet was extended. Concomitant mitral valvuloplasty (MVP) was performed in 4 patients. The competency of the aortic valve after completion of repair was evaluated by the transesophageal echocardiography in operating theater, and there was no aortic and mitral stenosis or regurgitation. In an early postoperative echocardiography, trivial AR was detected in 3 patients and mild MR in 1 patient. The end-systolic and end-diastolic dimensions of the left ventricle were decreased significantly(p<0.05) as compared with those of preoperative values. T ere was no mortality and no significant postoperative complication encountered. Late complication developed in 2 patients during the follow-up period(mean 7.9$\pm$ 5.9 months). One patient underwent AVR on postoperative 7th month due to endocarditis, and the another patient with Behcet's disease underwent Ross operation at postoperative 4th month. In conclusion, AVP of leaflet extension technique offers an excellent early clinical result and represents a good alterna!ivy surgical treatment for the pure AR especially in young age group, although long-term follow-up is necessary to determine the durability of glutaraldehyde-preserved autologous pericardium as a valve leaflet.

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Studies on the Post-hatching Development of Sertoli and Leydig Cells in the Testis of Korean Native Chickens (한국재래 닭 고환 지지세포와 간질세포의 부화 후 발달에 관한 연구)

  • Tae H. J.;Jang B. G.;Choi C. H.;Park Y. J.;Yang H. H.;Kim I. S.
    • Korean Journal of Poultry Science
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    • v.32 no.2
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    • pp.125-133
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    • 2005
  • Morphometric changes in testicular Sertoli and Leydig cells from hatching to adulthood were studied using Korean native chickens of 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 24, 28, 32, 44, 52 and 64 weeks (n=13 chickens per group) of age. The objective of this study was to understand the developmental phase of the Sertoli and Leydig cells with age. Testis of chickens was fixed by whole body perfusion using a fixative containing $2.5\%$ glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using 1 Um sections stained with methylene blue-azure II, qualitative and quantitative (stereological) morphological studies were performed. The average volume of a testis of 1 week old Korean native chickens was determined as $0.148\;cm^3$ and the parameter increased linearly from 1 week to 21 weeks days $(28.86\;cm^3)$, and did not change from 21 weeks to 64 weeks. The volume density of the seminiferous tubules increased with age from $32.6\%$ at week 1 to $92.89\%$ at week 64. The volume density of the interstitium represents $67.4\%$ of the testicular parenchyma at week 1. This proportion progressively diminished during development to reach a value of $7.11\%$ at week 64. The volume density of the Leydig cells decreased almost linearly from 1 week $(4.9\%)$ to 14 weeks $(1.7\%)$ and remained unchanged thereafter. In contrast, the Sertoli cells occupied a volume density of $3.4\%$ at week 1, increased progressively up to 18 weeks of age $(10.79\%)$ and remained unchanged thereafter. The absolute volume of the Leydig and Sertoli cells per testis increased significantly from week 1 to week 21 but did not change significantly from week 24 to week 64. The number of Leydig cells per testis increased almost linearly from 1 week to 21 weeks, remained high and unchanged with advancing age. The number of Sertoli cells per testis increased gradually with age from 1 week to 14 weeks and remained unchanged thereafter.

Ultrastructure of Degenerating Axon Terminals in the Basal Forebrain Nuclei of the Rat following Prefrontal Decortication (이마앞겉질을 제거시킨 흰쥐 앞뇌의 바닥핵무리에서 변성축삭종말의 미세구조연구)

  • Ahn, Byung-June;Ko, Jeong-Sik;Ahn, E-Tay
    • Applied Microscopy
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    • v.35 no.3
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    • pp.135-152
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    • 2005
  • Prefrontal cortex is a psychological and metaphysical cortex, which deals with feeling, memory, planning, attention, personality, etc. And it also integrates above-mentioned events with motor control and locomotor activities. Prefrontal cortex works as a highest CNS center, since the above mentioned functions are very important for one's successful life, and further more they are upgraded every moments through memory and learning. Many of these highest functions are supposed to be generated via forebrain basal nuclei (caudate nucleus, fundus striati nucleus, accumbens septi nucleus, septal nucleus, etc.). In this experiment, prefrontal efferent terminals within basal forebrain nuclei were ultrastructurally studied. Spraque Dawley rats, weighing $250{\sim}300g$ each, were anesthetized and their heads were fixed on the stereotaxic apparatus (experimental model, David Kopf Co.). Rats were incised their scalp, perforated a 3mm-wide hole on the right side of skull at the 11mm anterior point from the frontal O point (Ref. 13, Fig. 1), suctioned out the prefrontal cortex including cortex of the frontal pole, with suction instrument. Two days following the operations, small tissue blocks of basal forebrain nuclei were punched out, fixed in 1% glutaraldehyde-1% paraformaldehyde solution followed by 2% osmium tetroxide solutions. Ultrathin sections were stained with 1% borax-toluidin blue solution, and the stained sections were obserbed with an electron microscope. Degenerating axon terminals were found within all the basal forbrain nuclei. Numbers of degenerated terminals were largest in the caudate nucleus, next in order, in the fundus striati nucleus, in the accumbens septi nucleus, and the least in the septal nucleus. Only axospinous terminals were degenerated within the caudate nucleus and the fundus striati nucleus, and they showed the characters of striatal motor control system. Axodendritic and axospinous terminals were degenerated within the accumbens septi nucleus and the lateral septal nucleus, and they showed the characters of visceral limbic system. Prefrontal role in integrating the limbic system with the striatal system, en route basal forebrain nuclei, was discussed.

Histochemical Detection of Ionic Zinc in the Rat Olfactory Mucosa: Zinc Selenium Autometallography ($ZnSe^{AMG}$) (랫드 후각점막내 Zinc 이온의 조직화학적 동정)

  • Nam, Dong-Woo;Sun, Yuan-Jie;Kim, Sung-Joo;Kim, Yong-Kuk;Kim, Soo-Jin;Yu, Yun-Cho;Jeong, Young-Gil;Jo, Seung-Mook
    • Applied Microscopy
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    • v.33 no.2
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    • pp.145-154
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    • 2003
  • The present study was designed to demonstrate ionic zinc in the rat nasal mucosa by means of zinc selenium autometallography ($ZnSe^{AMG}$). Rats were given sodium selenide either intraperitoneally (i.p) or intranasally (i.n). Prior to the i.n. administration the rats were anesthetized with pentobarbital sodium (30 mg/kg, i.p.). A thin plastic tube coupled to a Hamilton syringe was then inserted into the right nostril and $10{\mu}l$ of the solution was instilled. For the i.p. administration non-anesthetized rats were given $100{\mu}l$ of the sodium selenide solution (10 mg/kg). Control rats were instilled with saline. After 2 hrs survival, the rats were anaesthetized and transcardially perfused with 3% glutaraldehyde. The olfactory area was removed and put into same fixative. The nose was then sectioned ($30{\mu}m$) horizontally, autometallography (AMG) was performed according to Danscher et al. (1997). After silver enhancement, fine AMG grains were scattered in the whole length of the olfactory epithelium containing olfactory receptor neurons, sustentacular and basal cells. However, much higher concentration of the AMG grains occupied near the surface and in the basal region of the olfactory epithelium. Both groups of i.p. and i.n. administration showed almost same level in the concentration of the AMG grains. In i.n. group, few AMG grains were also found in olfactory nerves of the lamina propria, suggesting zinc transport into the olfactory bulb via olfactory axons. At the electron microscopic level, the AMG grains were most entirely found in the supporting cells of the olfactory epithelium, and they were mostly localized in lysosome-like organelles. The i.n. group showed various signs of tissue damage of the olfactory mucosa, where dense concentration of AMG grains were localized at crystalloid structures. The present study demonstrated dense population of ionic zinc in the rat olfactory epithelium. zinc may play a role in the olfactory functioin and in the pathogenesis of the neurodegerative disorders affecting nose.