bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immunoelectron microscopy on lysis of the oocyte vitelline coat induced by a protein from spermatozoa in a marine mollusk, Tegula pfeifferi
Authors: Usui Noriko , Haino-Fukushima Kazu 
Published: June 2020

A protein responsible for lysis of the oocyte vitelline coat (VC), that is, a VC lysin, has been isolated from spermatozoa of the mollusk, Tegula pfeifferi. Biochemical studies using purified VC lysin and isolated VCs have revealed that the action of this VC lysin is stoichiometric rather than enzymatic. The present immunoelectron microscopic study was undertaken to obtain morphological evidence for this uncommon lytic mechanism using antibodies against the lysin and the soluble product (SP) released from the VC upon lysin treatment.Oocytes isolated from dissected ovaries were treated with the lysin (1 mg/ml) at 25°C for 10 min until their VCs were elevated high from the oocyte surface. The lysin-treated oocytes were prefixed with either 1% glutaraldehyde (GA) for cytochemical study or 2.5% GA plus tannic acid (TA) for morphological study. To localize the lysin and the SP in the VCs, prefixed oocytes were treated with ovalbumin followed by one of the antisera or preimmune serum (control).
A New Technique to Reveal DNA “En Bloc” and in Lowicryl Ultrathin Sections: The NaOH-Formaldehyde-MA-Method
Authors: Risueño M.C. , Testillano P.S. , Tandler C.J. , Sánchez-Pina M.A. , Olmedilla A. , Ollacarizqueta M.A. 
Published: June 2020

This paper deals with the developing of a new simple and repetitive technique to preferentially stain DNA-containing structures at the E.M. level that can be used “en bloc” and on sections. DNA staining techniques are scarce, being the osmium ammine one the only cytochemical method known until now that specifically stains DNA but not the proteins that are complexed with it, having been used to study chromatin ultrastructure. The osmium ammine synthesis is a complex process and, although an improvement has been lately reported, the stain is not comercially available.Glutaraldehyde-fixedAllium cepa L. root meristematic cells,Capsicum annuum L. & Scilla peruviana L. pollen grains were treated with 0.5N NaOH in 4% formaldehyde overnight, rinsed in water and 1% acetic acid, methanol dehydrated, and inmersed in methanol-acetic anhydride (5:1) at 25ºC overnight. Then, the samples were Epon embedded. To perform the technique on sections, some samples were glutaraldehyde or formaldehyde fixed, methanol dehydrated, Lowicryl embedded at -20ºC and ultrathin sections were treated in the same way as in the “en bloc” procedure.
Correlative Confocal and Electron Microscopy in Neurobiology
Authors: Deitch J.S. , Szarowski D.H. , Smith K.L. , Swann J.W. , Turner J.N. 
Published: June 2020

We have previously demonstrated the advantages of using confocal scanning laser microscopy (CSLM) in the reflection mode to image neurons labeled with the horseradish peroxidase (HRP) -- diaminobenzidine (DAB) method. Neurons which were impaled with microelectrodes could be characterized electrophysiologically, filled with horseradish peroxidase (or biocytin), reacted with DAB and then imaged in thick section. We have extended this technique to include electron microscopic observations. The optical sections generated with the CSLM helped in localizing specific labeled parts of the tissue that otherwise would require many hours of hunting in order to find.Neurons in 500-μm thick slices of rat hippocampus were filled with HRP via a recording micropipette and fixed for 4 h in 4% paraformaldehyde and 2.5% glutaraldehyde in Ringer’s solution. Vibratome sections were reacted with DAB (1.0 mg/ml) in 0.04% nickel ammonium sulfate, and placed in 1% OsO4 and then in uranyl acetate, each for 1 hour.
Laser Scanning Confocal Microscopy and Three-Dimesnsional Reconstruction of the Mitotic Spindles of Unequally Dividing Embryonic Cells
Authors: Holy J. , Schatten G. 
Published: June 2020

One of the classic limitations of light microscopy has been the fact that three dimensional biological events could only be visualized in two dimensions. Recently, this shortcoming has been overcome by combining the technologies of laser scanning confocal microscopy (LSCM) and computer processing of microscopical data by volume rendering methods. We have employed these techniques to examine morphogenetic events characterizing early development of sea urchin embryos. Specifically, the fourth cleavage division was examined because it is at this point that the first morphological signs of cell differentiation appear, manifested in the production of macromeres and micromeres by unequally dividing vegetal blastomeres.The mitotic spindle within vegetal blastomeres undergoing unequal cleavage are highly polarized and develop specialized, flattened asters toward the micromere pole. In order to reconstruct the three-dimensional features of these spindles, both isolated spindles and intact, extracted embryos were fluorescently labeled with antibodies directed against either centrosomes or tubulin.
Ultrastructural Cytochemical Study of Human Gastric Cancer: Observation of AKP ase,ACP ase,G6P ase,TPP ase and CO ase
Authors: Yuming Dong , Guanglin Yang , Wei Dong Du , Ai Liam Xu 
Published: June 2020

The activities and distributions of AKPase ,ACPase,G6Pase,TPPase and COase in human normal gastric mucosa and gastric cancer tissues were studied histochemically at light microscopic level. These enzymes are the marker enzymes of cell membrane lysosome endoplasmic reticulum, Golgi apparatus and mitochondrion objectively. On the basis of the research we set up a special ultrastructural cytochemical technique and first researched into gastric cancer domesticly. Ultrastructural cytochemistry is also called electron microscopic cytochemistry. This new technique possesses both the sensitivity of cytochemical reaction andi the high resolution of electron microscope. It is characterized by direct observation,exact localization and the combination morphology with function.The distributions of AKPase,ACPase,G6Pase,TPPase and COase in 14 cases of gastric cancer and 1 case of gastric Denign lesion were studied ultrastructurally. The results showed: 1. normal gastric epithelium had no AKPase reaction. The reaction of ACPase,G6Pase,TPPase and Coase were found in the corresponding organella, which were consistent with their function.
Ultrastructural Classification of Pituary Adenomas
Authors: Arumugam S. , Bharati Arumugam Sarasa 
Published: June 2020

Adenoaas of the pituitary are no longer classified based on their tinctorial affinity to dyes. With the advent of the newer methods of sophisticated technology, it is now possible to classify. These depending upon the type of hormone secreted based either on histochemical techniques or on ultrastructural characteristics. The latter provides an insight into the cytoplasmic organelle morphology which offers a delightful feast to the eye as well.This paper presents the ultrastructural characters of the pituitary adenoma as seen in Madras. 171 adenomas (124 males and 47 females) were seen during 1972-1989, classified at the light microscope level as 159 chromophobe, 2 basophilic, 4 eosinophilic and 6 mixed adenomas.Ultrastructural examination showed that the sparsely granular prolactin cell adenoma is the commonest adenoma to be encountered closely followed by the growth hormone cell adenoma, null cell adenoma, the mixed cell adenoma and others.
Cytological changes induced by platinum-thymine in sarcoma-180 cells: Electron microscopy study
Authors: Ofosu Gustav 
Published: June 2020

Platinum-thymine has been found to be a potent antitumor agent, which is quite soluble in water, and lack nephrotoxicity as the dose-limiting factor. The drug has been shown to interact with DNA and inhibits DNA, RNA and protein synthesis in mammalian cells in vitro. This investigation was undertaken to elucidate the cytotoxic effects of piatinum-thymine on sarcoma-180 cells in vitro ultrastructurally, Sarcoma-180 tumor bearing mice were treated with intraperitoneal injection of platinum-thymine 40mg/kg. A concentration of 60μg/ml dose of platinum-thymine was used in in vitro experiments. Treatments were at varying time intervals of 3, 7 and 21 days for in vivo experiments, and 30, 60 and 120 min., 6, 12, and 24th in vitro. Controls were not treated with platinum-thymine.Electron microscopic analyses of the treated cells in vivo and in vitro showed drastic cytotoxic effect.
Ultrastructural evaluation of chlorine inhalation injury following treatment with nebulized sodium bicarbonate
Authors: Langlinais P.C. , Okerberg C.V. , Chisholm C. , Kim S.H. , Mason A.D. , Pruitt B.A. 
Published: June 2020

Inhalation of chlorine gas is a common form of poisoning associated with industrial, household and transportation accidents. The use of inhaled 5% sodium bicarbonate solution for symptomatic treatment of chlorine gas injury has been suggested. Others have suggested possible intrapulmonary exothermic reaction from this procedure. The purpose of this study was to assess morphological alterations of the respiratory tract produced by inhalation of chlorine gas and to evaluate the therapeutic efficacy of inhaled nebulized 5% sodium bicarbonate after chlorine-induced injury.Twelve adult male random source sheep were conditioned two weeks prior to experimental use. The sheep were exposed to chlorine gas at the rate of 10 breaths/minute for 4 minutes. Immediately following chlorine exposure the sheep were placed on a respirator. Five minutes of nebulization with either 5% sodium bicarbonate or saline was done 30 minutes post-chlorine exposure. Necropsies were performed at 24 hours and tissue samples from two areas of the trachea and 8 areas of the lung were routinely processed for light and transmission electron microscopy.
Histopathological observations of recurrent experimental autoimmune encephalomyelitis (rEAE) in female Lewis rats
Authors: Aschenbrenner J. E. , Jackson F. L. , Orr E. L. , Chang I. F. , Oakford L. X. 
Published: June 2020

Experimental autoimmune enchephalomyelltis (EAE) is a disease which can be induced in Lewis rats by exposure of the animal to a source of myelin in the presence of adjuvant. Similar symptoms and histopathology are observed In people with multiple sclerosis (MS) leading to the widespread use ofEAEas an animal model forMS (1). Induction ofEAEin the male Lewis rat results In a monophasic disease characterized by a single severe occurrence of symptoms followed by spontaneous recovery. Female Lewis rats when exposed to identical stimuli will develop a recurrent form of the disease (rEAE) characterized by a temporally consistent, spontaneous secondary occurrence of clinical symptoms(2), which more closely mimics humanMS.The goal of this preliminary study was to characterize the disease process in female Lewis rats (rEAE) using both light and transmission electron microscopy (TEM). Samples of lumbrosacral spinal cord were chosen for Initial examination. This tissue was sampled at four stages of the disease process, namely: in the primary occurrence phase (Peak 1, Day 12), the first recovery phase (Day 18), the secondary occurrence (Peak 2, Day 24) and the second recovery phase (Day 31) (seeFigure 1). Lumbrosacral spinal cord tissue from non-sensitized female Lewis rats, were used as control tissue.
Ultrastructural study of the Pseudopregnant Rat Uterus treated with Antiprogestins
Authors: Singh G. , Maitra S.C. , Singh M.M. 
Published: June 2020

Steroidal antiprogestins RU-38486 and ZK-98734 have been reported to prevent progesterone induced uterine endometrial proliferation in estrogen primed rabbits, block decidual response and implantation in rats and induce abortion in rats, rabbits, monkeys and humans.The present study is aimed to generate information about the effect of these antigestagens on uterine morphology and ultrastructure in the pseudopregnant rats.Unilateral intraluminal peanut oil instillation on day 5 post-coitum in pseudopregnant rats induced marked decidualization as characterized by 560% increase in uterine weight. Histologically, it showed narrow lumen, low cuboidal epithelium with large rounded nuclei, loose stroma, large decidual cells (Fig. 1) and glands with closed lumen (Fig. 5). The epithelial surface appeared polygonal with short, stubby and sparse microvilli (Fig. 2), ultrastructurally, luminal as well as glandular epithelium had abundant mitochondria, polyribosomes, secretory granules, active golgi, and numerous pinocytotic vesicles representing functionally active status (Figs. 7 & 11). Decidual cells had numerous mitochrondia, inflated RER and irregular surface infoldings (Fig. 9).
vitreal cells and specialized phagocytes in the chick embryo retina: A TEM and SEM study
Authors: Cuadros M.A. , Martinez-Guerrero M.J. , Rios A. 
Published: June 2020

In the chick embryo retina (days 3-4 of incubation), coinciding with an increase in cell death, specialized phagocytes characterized by intense acid phosphatase activity have been described. In these preparations, all free cells in the vitreal humor (vitreal cells) were strongly labeled. Conventional TEM and SEM techniques were used to characterize them and attempt to determine their relationship with retinal phagocytes.Two types of vitreal cells were distinguished. The first are located at some distance from the basement membrane of the neuroepithelium, and are rounded, with numerous vacuoles and thin cytoplasmic prolongations. Images of exo- and or endocytosis were frequent; the cells showed a well-developed Golgi apparatus (Fig. 1) In SEM images, the cells was covered with short cellular processes (Fig. 3). Cells lying parallel to or alongside the basement membrane are elongated. The plasma membrane is frequently in intimate contact with the basement membrane. These cells have generally a large cytoplasmic expansion (Fig. 5).
Elemental composition of mast cell granules of the rat and tree frog: X-ray Microanalysis on fresh frozen dried ultrathin sections
Authors: Takaya Kenichi 
Published: June 2020

Electron probe x-ray microanalysis using fresh air-dried spreads revealed electrolyte elements in the granules of platelets, mast cells, pancreatic acinar cells and melanocytes. Mast cell granules of the rat and tree frog are quantitatively compared by an energy dispersive spectrometry (EDS) using fresh frozen dried ultrathin sections.Adult Wistar rats (ca. 250 g) and tree frogs (Hyla arborea japonica) of both sexes were used. Fresh frozen dried smears of rat peritoneal mast cells were prepared on the collodion-membrane covered titanium grid. Fresh frozen dried ultrathin sections of the tongue of the rat and tree frog were made by the metal contact method employing the rapid freezing apparatus (RF-2) cooled with liquid nitrogen. Ultrathin cryosections (60 nmj were cut at 163 K, transferred to the frozen specimen treating apparatus (FD-2) for freeze-drying in high vacuum at 173 K for 74-96 h. The specimens were observed first by 200kV transmission electron microscopy (TEM) and then under the scanning analytical electron microscope (X-650) for scanning transmission electron microscopy (STEM) images at an acceleration voltage of 40 kV and a specimen current of 0.2 nA.
Endocytosis and intracellular transport of colloidal gold-labeled ConA in mouse peritoneal macrophage
Authors: Piao Yingjie , Zhou Yimo , HU Wen 
Published: June 2020

The purpose of this study is to investigate the endocytosis and intracellular transport of colloidal gold-labeled ConA (CG-ConA) in mouse peritoneal macrophages using pre-embedding staining method.Labeling with CG-ConA, acid phosphotase (AcPase) cytochemistry and electron microscope (EM). Mouse peritoneal macrophages were elicited by intraperitoneal injection of glycogen, after which 1ml CG-ConA (CG 1mg/ml) was injected per mouse. At different intervals after injection of CG-ConA (5,10,20 and 30 min), the peritoneal macrophages were isolated and treated for their AcPase activity according to the produce of Gomori. The CG-ConA and AcPase activity in these macrophages were examined under TEM.Surface distribution of CG-ConA 5-1Omin after intraperitoneal injection of CG-ConA many clusters and clumps of CG-ConA particles were formed on the macrophages (Fig. 1). The cluster formation is temperature-dependent and receptor-mediated, because the same phenomenon could be observed in the elicited macrophages after incubation with CG-ConA at 37°C in vitro, but not in those at 0-4°C and those incubated with pure CG particles. At 0-4°C CG-ConA particles were randomly and diffusely distributed over the cell surface. Aggregates of CG-ConA particles were often found between cellular processes.
Ultrastructural investigations of MDL 19,660-induced effects on the canine platelet and megakaryocyte
Authors: Loudy D.E. , Sprinkle-Cavallo J. , Yarrington J.T. , Thompson F.Y. , Gibson J.P. 
Published: June 2020

Previous short term toxicological studies of one to two weeks duration have demonstrated that MDL 19,660 (5-(4-chlorophenyl)-2,4-dihydro-2,4-dimethyl-3Hl, 2,4-triazole-3-thione), an antidepressant drug, causes a dose-related thrombocytopenia in dogs. Platelet counts started to decline after two days of dosing with 30 mg/kg/day and continued to decrease to their lowest levels by 5-7 days. The loss in platelets was primarily of the small discoid subpopulation. In vitro studies have also indicated that MDL 19,660: does not spontaneously aggregate canine platelets and has moderate antiaggregating properties by inhibiting ADP-induced aggregation. The objectives of the present investigation of MDL 19,660 were to evaluate ultrastructurally long term effects on platelet internal architecture and changes in subpopulations of platelets and megakaryocytes.Nine male and nine female beagle dogs were divided equally into three groups and were administered orally 0, 15, or 30 mg/kg/day of MDL 19,660 for three months. Compared to a control platelet range of 353,000- 452,000/μl, a doserelated thrombocytopenia reached a maximum severity of an average of 135,000/μl for the 15 mg/kg/day dogs after two weeks and 81,000/μl for the 30 mg/kg/day dogs after one week.
High-voltage electron microscopy and 3-D reconstruction of solitary chemosensory cells and Di-I labeling of primary afferent nerves
Authors: Kotrschal Kurt , Kinnamon John C. , Royer Suzanne M. 
Published: June 2020

Solitary chemosensory cells (SCCs) resembling taste bud cells in shape and ultrastructure have been found in fish and amphibians. A quantitative survey in teleosts has revealed that SCCs may be even more abundant than taste bud cells. Due to their scattered distribution, however, this abundant and potentially important vertebrate chemosensory system has eluded closer examination. This situation has changed with the introduction of a suitable research model: the anterior dorsal fin (ADF) of rocklings (Gadidae, Teleostei). This specialized chemosensory organ contains several million SCCs that converge onto fine caliber recurrent facial nerve fibers. The ADF model has allowed us to investigate SCC innervation, fine structure and function.Using high voltage electron microscopy of serial sections and computer-assisted three-dimensional reconstructions, we have obtained significant quantitative data on the spatial structure of SCC assemblages in the ADF epidermis and of synaptic contacts between SCCs and innervating facial nerves. Approximately 15% of all cells present in the ADF epidermis are SCCs, making up 30% of the epidermal volume. Unlike taste buds, the ADF contains only a single sensory cell type and lacks basal cells or specialized “supporting cells.”
Application of stereological analysis of cell volume to isolated myocytes in culture with and without adrenergic innervation
Authors: Rosenthal Jane K. , Atkins Dianne L. , Marvin William J. , Krumm Penny A. 
Published: June 2020

To comprehend structural changes in cardiac myocytes accompanying adrenergic innervation, it is essential that a three dimensional analysis be performed. To date, biological studies which utilize stereological methods have been limited to cells in tissue and in organs. Our laboratory has utilized current stereological techniques for measuring absolute volumes of individual myocytes in primary culture. Cell volumes are calculated for two distinct groups of cells at 96 hours in culture: isolated myocytes and myocytes innervated with adrenergic neurons (Figure 1).Cardiac myocytes are cultured from the ventricular apices of newborn rats. Cells are plated directly onto tissue culture dishes with or without preplated explants from the paravertebral thoracolumbar sympathetic chain. On day four cultures are photographed and marked for one-to-one cell location. Following conventional fixation and embeddment in eponate-12, the cells are relocated and mounted for microtomy. The cells are completely sectioned at 120nm in their parallel orientation to the surface of the dish (Figure 2). Serial sections are collected on formvar coated slotted grids and are recorded in sequence.
The cytoplasmic surface of acetylcholine receptor enriched Xenopus myocytes
Authors: Samuelsson Steven J. 
Published: June 2020

During neuromuscular synaptogenesis nicotinic acetylcholine receptors (nAChR) diffusely arranged in the postsynaptic membrane preferentially sequester into aggregates. With time these aggregates come to collocalize with the post-junctional folds where they function as neurotransmitter receptors. I am interested in the molecular mechanism which maintains this integral membrane protein in place.I present a high resolution account of the muscle membrane/cytoskeleton at the, nAChR plaques.Cocultures of myocytes and neural tube neurons from stage 22 Xenopus laevis embryos develop a reasonable number of ventral surface junctions. nAChRs also aggregate in muscle cultures without neurons. Receptor plaques are identified by labeling with fluorescent alpha-bungarotoxin (BTX)(Fig. 1a, 2a). In order to gain access to the cytoplasmic surface the cultures are incubated in a ZnCl2 solution, which stabilizes the R-rich ventral membrane areas, then sheared with a stream of ice-cold buffer. After immediate fixation the cellular fragments are ready for ultrastructural processingor immunocytochemistry.The quick-freeze, deep-etch, rotary-shadow technique is best suited for high-resolution en-face observation of these preparations. Improvements in the original technique were necessary to produce a satisfactory and informative replica.
Ultrastructural study on the development of rat amygdaloid body
Authors: Manolova A. , Manolov S. 
Published: June 2020

Relatively few data on the development of the amygdaloid complex are available only at the light microscopic level (1-3). The existence of just general morphological criteria requires the performance of other investigations in particular ultrastructural in order to obtain new and more detailed information about the changes in the amygdaloid complex during development.The prenatal and postnatal development of rat amygdaloid complex beginning from the 12th embrionic day (ED) till the 33rd postnatal day (PD) has been studied. During the early stages of neurogenesis (12ED), the nerve cells were observed to be closely packed, small-sized, with oval shape. A thin ring of cytoplasm surrounded their large nuclei, their nucleoli being very active with various size and form (Fig.1). Some cells possessed more abundant cytoplasm. The perikarya were extremely rich in free ribosomes. Single sacs of the rough endoplasmic reticulum and mitochondria were observed among them. The mitochondria were with light matrix and possessed few cristae. Neural processes were viewed to sprout from some nerve cells (Fig.2). Later the nuclei were still comparatively large and with various shape.