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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Korean Society of Electron Microscopy
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Volume 40, Issue 4 - Dec 2010
Volume 40, Issue 3 - Sep 2010
Volume 40, Issue 2 - Jun 2010
Volume 40, Issue 1 - Mar 2010
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Applications of Focused Ion Beam for Biomedical Research
Kim, Ki-Woo ; Baek, Saeng-Geul ; Park, Byung-Joon ; Kim, Hyun-Wook ; Rhyu, Im-Joo ;
Applied Microscopy, volume 40, issue 4, 2010, Pages 177~183
A focused ion beam (FIB) system produces a beam of positive ions (usually gallium) which are heavier than electrons and can be focused by electrostatic lenses into a spot on the specimen. With its ability milling of the specimen material by 10 to 100 nm with each pass of the beam, FIB is widely adopted in materials science, semiconductor industry, and ceramics research. Recently, FIB has been increasingly employed in the field of biomedical sciences. Here we provide a brief introduction to FIB and its applications for a wide variety of biomedical research. The surface of specimen can be in situ processed and quasi-real time visualized by two beam combination of FIB and field emission scanning electron microscope (FESEM). Due to its milling process, internal structures can be exposed and analyzed: yeast cells, fungus-inoculated wheat leaf, mannitol particles in inhalation aerosols, and oyster shell. Serial blockface tomography with the system kindles 3-dimensional reconstruction researches in the realm of nervous system and life sciences. Two-beam system of FIB/FESEM is a versatile tool to be utilized in the biomedical sciences, especially in 3-dimensional reconstruction studies.
Autoradiographic Verification of Transdermal Penetration of Oleic Acid-conjugated Peptide Nanosomes
Lee, Kyung-Eun ; Jung, Min-Kyo ; Eum, Jai-Hoon ; Jung, Se-Hui ; Ha, Kwon-Soo ; Park, Jeong-Hae ; Lee, Jin-Sung ; Han, Sung-Sik ; Choe, Myeon ;
Applied Microscopy, volume 40, issue 4, 2010, Pages 185~191
Short peptides are potentially effective materials as cosmeceuticals, but their delivery across the skin can be problematic due to the ionic nature of peptides and the structure of the skin. For short peptide to be utilized as cosmeceuticals, its ability to penetrate the skin must be altered. In this study, we conjugated the widely used procollagen type I signal peptide, KTTKS, with oleic acid to improve the lipophilic properties of the peptide, and used the oleic acid-conjugated peptides to construct cosmeceutical nanosomes. Then we examined the penetration of cosmeceutical nanosomes prepared from isotope-labeled peptide into the skin after transdermal application using autoradiography. Because of its hydrophilic property of penta-peptide, the penta-peptide itself was not able to be penetrated through the stratum corneum of the skin. In contrast, nanosomes made from olecic acid conjugated penta-peptide were able to be penetrated through the stratum corneum effectively. Autoradiography showed the precise penetration points to dermal layer, demonstrating the appropriateness of this method for clarifying the mechanism of penetration of transdermal delivery systems.
Effects of Bacillus Calmette-Guerin (BCG) on the DNA Synthesis of Mouse Duodenal Mucosal Epithelial Cells Inoculated with Ehrlich Carcinoma Cells
Ko, Jeong-Sik ; Kim, Heung-No ; Park, Kyung-Ho ; Park, Dae-Kyoon ; Kim, Duk-Soo ;
Applied Microscopy, volume 40, issue 4, 2010, Pages 193~200
This experiment was performed to evaluate the morphological responses of the duodenal epithelial cells of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of Bacillus Calmette-Guerin (BCG). In the experimental groups, each mouse was inoculated with
Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.2 mL of saline or BCG (0.5 mL/25 g B.W.:
CFU) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of saline or BCG, each mouse was injected with a single dose of
/g of methyl-
-thymidine (25 Ci/mmol, Amersham Lab, England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed, and duodenal tissues were taken and fixed in 10% neutral formalin. Deparaffinized sections were coated with autoradiographic emulsion EM-1 (Amersham Lab, England) in a dark room and dried and were placed in a light-tight box. The number of labeled epithelial cells in the duodenal mucosae (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. On the light microscopic study, duodenal mucosae had no severe morphological changes following the injection of BCG. In the tumor control and BCG treated groups, a number of small lymphocytes and eosinophile leucocytes are slightly increased as compared with those of the normal control ones. On the autoradiographic study, number of the labeled cells of normal control, tumor control and BCG-treated mice were 632.3 (
), 761.7 (
) and 505.0 (
) respectively. From the above results, BCG may suppress the DNA synthesis of the duodenal epithelial cells, but does not results severe structural defect on the duodenal mucosae. And it is suggested that BCG may greatly improve the anticancer therapy on certain kind of cancer.
Inhibitory Effects of Several Korean Traditional Herbs on Anaphylactic Reaction and Mast Cell Activation
Chai, Ok-Hee ;
Applied Microscopy, volume 40, issue 4, 2010, Pages 201~209
Korean traditional herbs, especially Anemarrhena asphodeloides (A. asphodeloides), Salvia miltiorrhiza (S. miltiorrhiza), and Terminalia chebula (T. chebula) have been known to have the immunomodulatory, anti-tumor, and anti-inflammatory effects. However, direct cellular mechanism underlying the mast cell-mediated anaphylaxis-like reaction has poorly been understood. In the present study, the effects of the methanol extracts of A. asphodeloides (MEAA), S. miltiorrhiza (MESM), and T. chebula (METC) on anaphylactic reaction were investigated. Using in vitro and in vivo experiments, the influences of MEAA, MESM, and METC on the compound 48/80-induced anaphylaxis-like reaction and mast cell activation, and IgEmediated PCA were examined. Results are below; 1) MEAA, MESM, and METC significantly inhibited systemic anaphylaxis-like reaction, ear swelling response, and IgE-mediated PCA. 2) the compound 48/80-induced mast cell degranulation, histamine release of rat peritoneal mast cells (RPMC) were significantly inhibited by the pretreatment with MEAA, MESM, and METC, and 3) the compound 48/80-induced calcium influx in RPMC was significantly inhibited by the pretreatment with MEAA, MESM, and METC. These results suggest that MEAA, MESM, and METC may be an activity to inhibit the compound 48/80-induced or anti-DNP IgE-mediated anaphylactic reactions by blocking histamine release and calcium uptake into RPMC. MEAA, MESM, and METC potentially may serve as an effective therapeutic tool for allergic diseases.
The Oxytocinergic Neurons in Hypothamo-hypophysial Tract Contributes to CNS Pathway Innervating Ovary in Rat
Byun, Kyung-Hee ; Oh, Jee-Hyun ; Jo, Seung-Mook ; Lee, Bong-Hee ;
Applied Microscopy, volume 40, issue 4, 2010, Pages 211~218
The mammalian ovary is innervated by sympathetic and sensory neurons which contribute to regulating several aspects of ovarian function, including blood flow, steroidogenesis and follicular development. The existence of a neural connection between central neurons and the ovary has been rarely reported, but the mechanism underlying integration of ovarian activity to broader neuroendocrine responses has not been reported. We have now used a viral transneuronal tracing technique combined with a conventional retrograde labeling procedure of CT-HRP to demonstrate that oxytocin-producing neurons of the hypothalamus are synaptically connected to the ovary. Since ovarian activity is suppressed but the activity of oxytocin neurons is increased during breast feeding. Our finding that the oxytocinergic neural connection is likely to provide a direct transsynaptic mechanism by which the central nervous system maintains the state of infertility that accompanies lactation in mammals.
Seed Morphology of Euphorbia Section Anthacantha (Euphorbiaceae) and Related Taxa
Na, Hee-Jung ; Park, Ki-Ryong ;
Applied Microscopy, volume 40, issue 4, 2010, Pages 219~228
The seeds of 16 species from sect. Anthacantha and related taxa were examined using light and scanning electron microscopy to illucidate the boundary of the section and their relationship among species. Using six qualitative characters clustering analyses were conducted, and three types were recognized. Type I including species from sect. Anthacantha+sect. Meleuphorbia is characterized by the ovate shape, rounded at base and smooth along the ventral line. Type II including species from sect. Medusae+sect. Treisia-1 is squared in shape and are tuberculate. Tubercles are prominent along the ventral line. Type III (sect. Treisia-2 group) is mostly rounded, and as in type II, tubercles are prominent along the ventral line, but the unique crestae consisting of the grouping testa cells differed from the surface patterns of Types I and II. Based on the seed morphology, sects. Anthacantha and Meleuphorbia are closely related, which is well supported by the results from molecular and pollen morphological studies. Besides, sects. Treisia and Medusae are not closely related in terms of seed characters, and this is not consistent with the results of recent molecular studies.
Synergistic Effects of 5-Fluorouracil (FU) and Curcumin on Human Cervical Cancer Cells
Ahn, Seong-Ho ; Kim, Dong-Heui ; Kang, Jung-Hoon ; Lee, Myeong-Seon ;
Applied Microscopy, volume 40, issue 4, 2010, Pages 229~235
Cervical cancer is associated with low antioxidant status. It has a high prevalence especially amongst woman in Asia and is a leading cause of cancer death. Cancer chemotherapy in vivo improved in cases with high p53 expression in the tumor tissue. The restoration of p53 levels could be a potential strategy to increase chemoresponsiveness. Under circumstances where damage is extensive, p53 plays a direct role in trigering apoptosis. To investigate the effect of curcumin (CMN) as an antioxidant agent on anticancer agent 5-fluorouracil (5FU) induced apoptosis and p53 expression, HPV-18 positive HeLa cells were treated with noncytotoxic amounts of antioxidant. Curcumin induced apoptosis in cervical cancer cells. Morphological hallmarks of apoptosis such as nuclear fragmentation and internucleosomal fragmentation of DNA were observed. CMN caused upregulation of p53 expression, evident from Western blotting data and also increased the susceptibility/apoptosis induced by 5FU. These results show that increasing drug sensitivity of cervical cancer cells by upregulation of p53 using CMN is novel approach and could have a possible therapeutic potential in cervical cancer.
Morphological Differentiation of the Trap in Aquatic and Terrestrial Utricularia Species
Lee, Kyoung-Lan ; Kim, In-Sun ;
Applied Microscopy, volume 40, issue 4, 2010, Pages 237~244
Utricularia forms small, but complex carnivorous trap along the stem either in water or in soil depending upon species. The shapes and sizes of the traps, appendages, and trichomes are known to differ among aquatic, terrestrial and epiphytic species. In the present study, the morphology and microstructure of the trap in aquatic Utricularia japonica and terrestrial U. livida were examined using light and electron microscopy. The aim of this study was to compare the characteristics of trap features between the aquatic and terrestrial species. The trap was found to be comprised of a thin walled bladder with numerous capitate trichomes, two-armed bifid and four-armed quadrifid glands in both species; however, the traps of the two species were different in size, and number and morphology of the trichomes and glands. Aquatic Utricularia was chlorenchymatous with chloroplasts distributed throughout the body, whereas the terrestrial species was translucent without plastids due to an adaptation to underground habitats. Furthermore, the former differed considerably in that the traps developed antenna and appendages around the entrance area. A peculiar trap entrance was also noted in U. livida, which exhibited radiating rows of various trichomes within funnel-shaped tissue. A large number of glandular trichomes covered the entrance area and door surface with four trigger hairs each in the aquatic form but only two in the terrestrial form. The glandular trichomes near the door secreted a large amount of mucilage that temporarily composed the velum in the U. japonica, however, it was not observed in the terrestrial species. All of the aforementioned features were highly related in their structure and function during carnivorous mechanism in Utricularia. The current findings provide important data for further comparison of the different life forms within Utricularia.
Effect of Epidermal Changes in the Mice Skin Following Glycolic Acid Peeling
Lee, Jee-Yean ; Lee, Suk-Jun ; Chang, Byung-Soo ;
Applied Microscopy, volume 40, issue 4, 2010, Pages 245~251
In this study, we applied 15% glycolic acid peeling agent and 30% glycolic acid peeling agent to the dorsal skin of mice and analyzed the oil content, moisture content, and pH value of the skin before and after test using cutometer. Ultrastructure for changes in the epidermis of mice were observed under a dermoscope and a scanning electron microscope before and after test. When using 15% glycolic acid peeling agent, the changes of moisture content was measured 74.67 AU for the normal control group and 70.21 AU for the experimental group, the oil content was 13.49 mg/
for the normal control group before test and 8.25 mg/
after test, and the pH value was 6.70 and 5.36 before and after test, respectively. When using 30% glycolic acid peeling agent, the moisture content was measured 74.46 AU for the normal control group before test and 53.50 AU for the experimental group after test, the oil content was 13.82mg/
and 5.70 mg/
before and after test, and the pH value was 6.45 and 4.58 before and after test, respectively. As such, it was found that the changes of moisture and oil content on the skin rely on the concentration of peeling agent and the degree of exfoliation of keratin. The surface of stratum corneum of mice with application of 15% glycolic acid peeling agent was relatively smooth and the exposed cellular surface of keratinocyte had some wrinkles. The surface of stratum corneum of mice with application of 30% glycolic acid peeling agent was smooth. No wrinkles were observed under high-resolution scanning electron microscope.
Ultrastructure of the Sperm in Testes of the Pond Smelt (Hypomesus nipponensis)
Kim, Jae-Goo ; Park, No-Kwan ; Reu, Dong-Suck ;
Applied Microscopy, volume 40, issue 4, 2010, Pages 253~259
The ultrastructure of sperms in testes of the pond smelt (Hypomesus nipponensis) was investigated using electron microscopes. The whitish testis was located between swim bladder and intestine. Especially, the left testis was larger than the right testis. The sperm was approximately
in length. The sperm had an oval head and the acrosome was not found. The nucleus was about 400 nm in diameter and chromatin was incompletely condensed. The nuclear fossa deeply formed in sperm head and two centrioles were located in the fossa. The mitochondrium was observed only one in midpiece of the sperm and a motile flagellum consisted of an axoneme with a typical 9+2 pattern of microtubule. Also, the tail of the sperm has axonemal fins.
The Distribution of ATPase and Porin in the Bovine Heart Mitochondrial Cristae
Kim, Tae-Keun ; Min, Byoung-Hoon ; Kim, Soo-Jin ;
Applied Microscopy, volume 40, issue 4, 2010, Pages 261~266
ATP is the energy source synthesized at the electron transferase that consist of complex I, II, III, IV and V in mitochondrial cristae. The complex V functions as ATPase which composed of sub-complex
. Porin or VDAC (voltagedependent anion-selective channel), is a family of small pore-forming proteins of the mitochondrial outer membrane, and play important roles in the regulated flux of anion, proton and metabolites between the cytosolic and mitochondrial compartments. The channel allows the diffusion of negatively charged solutes such as succinate, malate, and ATP in the fully open state, but of positively charged ions in subconducting state. In this study, in order to investigate the relationship of the function and localization between porin and ATPase we observed the distribution of porin and ATPase in the mitochondria of the bovine heart. Monoclonal antibodies against porin and ATPase
-subunit were used to detect porin and ATPase using light microscope with immunohistochemistry and immunofluorescence, and using electron microscope with immunogold-labeling. ATPase were stained in longitudinal section region in cardiac muscle, porin were stained in longitudinal section region in cardiac muscle. We viewed more specific pattern of localization and distribution of these proteins using immunofluorescence method. There were some region which were labeled with porin or ATPase respectively, and others which were labeled both proteins in cardiac muscle. The electron microscope results showed that immunogold labeled porin were labeled locally at mitochondrial outer membrane and ATPase were labeled evenly at mitochondrial cristae. But ATPase was not labeled at mitochondria cristae. These results confirmed the subcellular localizations of porin and ATPase in mitochondrial outer membrane and cristae. Also, we assumed that ATP synthesis always does not activation in all mitochondria exist in the bovine cardiac muscle.
Scanning Electron Microscopic Observation of Human Skin Replica
Rhyu, Yeon-Seung ; Chung, Ye-Ji ; Uhm, Chang-Sub ;
Applied Microscopy, volume 40, issue 4, 2010, Pages 267~270
The skin is the largest organ of the integument system whose surface is closely related with many physiological and pathological conditions. Various methods are used to understand the structural and functional status of human skin. We would like to present usefulness of scanning electron microscopic (SEM) observation of skin replica and its significance of training module for a novice. The silicon replicas from several regions of the body (hand, finger, forearm, lip, and face) were casted by applying Exafine
mixture. The positive replicas were prepared by applying EPON 812 mixture on negative silicon replicas. Some of the negative silicon replicas were cut with a razor blade and surface profiles were observed. The negative and positive replicas were coated with platinum and were observed under the scanning electron microscope. We could investigate the detailed structures of the human skin surface without any physical damage to the subject. The positive replicas depicted real surface structure of the human skin vividly. The cross sectional view of the negative silicon replicas provided surface profile clearly. The scanning electron microscopic observation of the human skin replicas would be useful to study skin surface structures and to evaluate medical and esthetical applications.