Fine structure of human sperm entry into zona-free hamster oocyte

1978 ◽  
Vol 36 (2) ◽  
pp. 540-541
Author(s):  
C. Barros ◽  
J. González ◽  
E. Herrera ◽  
E. Bustos-Obregón
Keyword(s):  
Osmium Tetroxide ◽  
Human Sperm ◽  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Human Spermatozoa ◽  
Low Viscosity ◽  
Cacodylate Buffer ◽  
Ultrathin Sections ◽  
Hamster Oocyte

Zona-free mammalian oocytes have been observed to fuse, under in vitro conditions, With non-homologous spermatozoa. Taking advantage of this heterologous gamete fusion, we designed a bioassay to test -by means of zona-free hamster oocytes- the fertile ability of human spermatozoa, from semen samples of patients attending an Infertility Clinic. To further validate our bioassay, which Was reported elsewhere, we studied the behavior of gamete membranes during fusion at the ultrastructural level.Zona-free hamster oocytes were mixed in vitro with human spermatozoa. At different times after the start of incubation, oocytes were fixed in 1% glutaraldehyde in 0. 25M cacodylate buffer pH 7. A and post-fixed in 196 osmium tetroxide. After dehydration in acetone, they were embedded in a low viscosity epoxy resin. Ultrathin sections were stained with uranyl acetate and lead citrate and examined with a Phillips 300 electron microscope.

Myocardial Alterations in Animals Following High Altitude Exposure

1968 ◽  
Vol 26 ◽  
pp. 114-115
Author(s):  
M. B. Bischoff ◽  
W. D. Dean ◽  
T. J. Bucci ◽  
L . A. Frics
Keyword(s):  
Pulmonary Artery ◽  
Right Ventricular ◽  
High Altitude ◽  
Sea Level ◽  
Osmium Tetroxide ◽  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Heart Weight ◽  
Morphological Evidence ◽  
Ultrathin Sections

There is a paucity of morphological evidence, particularly at the ultrastructural level, to correlate with tile physiological effects of chronic exposure to high altitude. In this study the myocardium of dogs and rabbits subjected to five months exposure at an altitude of 14,110 feet was compared to that of animals residing at sea level. The animals kept at 14,110 feet were shipped from sea level (160 feet) witnout acclimatization at intermediate altitudes.The physiological parameters including the hematologic changes, pulmonary artery pressures, pulmonary artery oxygen saturations, electrocardiograph changes, right ventricular weight to body weight and right ventricular weight to total heart weight ratios observed in these animals have been reported previously.The animals were necropsied at the end of the five months. The myocardium was quickly removed, cut into small pieces and immersed in 1% osmium tetroxide buffered with veronal acetate. Following fixation the tissues were dehydrated in ascending ethanol concentrations followed by propylene oxide and embedded in Epon 812. Ultrathin sections were doubly stained in uranyl acetate and lead citrate.

Ultrastructural changes in sciatic nerve tissue: Effects of passive maternal smoking

1983 ◽  
Vol 41 ◽  
pp. 650-651
Author(s):  
Amankwah K.S. ◽  
A.D. Weberg ◽  
R.C. Kaufmann
Keyword(s):  
Sciatic Nerve ◽  
Maternal Smoking ◽  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Nerve Tissue ◽  
Ultrastructural Changes ◽  
Cacodylate Buffer ◽  
And Control ◽  
Spontaneous Delivery

Previous research has revealed that passive (involuntary inhalation) tobacco smoking during gestation can have adverse effects upon the developing fetus. These prior investigations did not concentrate on changes in fetal morphology. This study was undertaken to delineate fetal neural abnormalities at the ultrastructural level in mice pups exposed in utero to passive maternal smoking.Pregnant study animals, housed in a special chamber, were subjected to cigarette smoke daily from conception until delivery. Blood tests for determination of carbon monoxide levels were run at 15-18 days gestation. Sciatic nerve tissue from experimental and control animals were obtained following spontaneous delivery and fixed in 2.5% gluteraldehyde in 0.1M cacodylate buffer pH 7.3. The samples were post-fixed in osmium ferrocyanide (1:1 mixture of 1.5% aqueous OSO4 and 2.5% K4 Fe(CN)6). Following dehydration, the tissues were infiltrated with and embedded in Spurr. Sections were stained with uranyl acetate and lead citrate.

The Ultrastructure of Metastatic Human Mammary Carcinoma After Prolonged Estrogen Therapy

1967 ◽  
Vol 25 ◽  
pp. 180-181
Author(s):  
John H.L. Watson ◽  
John L. Swedo ◽  
R.W. Talley
Keyword(s):  
Mammary Carcinoma ◽  
Osmium Tetroxide ◽  
Estrogen Therapy ◽  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Human Mammary Carcinoma ◽  
Surgical Biopsy ◽  
Lead Hydroxide ◽  
Preliminary Study ◽  
Hormonal Therapies

A preliminary study of human mammary carcinoma on the ultrastructural level is reported for a metastatic, subcutaneous nodule, obtained as a surgical biopsy. The patient's tumor had responded favorably to a series of hormonal therapies, including androgens, estrogens, progestins, and corticoids for recurring nodules over eight years. The pertinent nodule was removed from the region of the gluteal maximus, two weeks following stilbestrol therapy. It was about 1.5 cms in diameter, and was located within the dermis. Pieces from it were fixed immediately in cold fixatives: phosphate buffered osmium tetroxide, glutaraldehyde, and paraformaldehyde. Embedment in each case was in Vestopal W. Contrasting was done with combinations of uranyl acetate and lead hydroxide.

Opening images of synaptic vesicles and calcium localization by means of microwave fixation method

1990 ◽  
Vol 48 (2) ◽  
pp. 182-183
Author(s):  
Vinci Mizuhira ◽  
Hiroshi Hasegawa
Keyword(s):  
Synaptic Vesicles ◽  
Room Temperature ◽  
Osmium Tetroxide ◽  
Sampling Procedure ◽  
Fixation Method ◽  
Potassium Oxalate ◽  
X Ray ◽  
Cacodylate Buffer ◽  
Ultrathin Sections ◽  
Microwave Fixation

Microwave irradiation (MWI) was applied to 0.3 to 1 cm3 blocks of rat central nervous system at 2.45 GHz/500W for about 20 sec in a fixative, at room temperature. Fixative composed of 2% paraformaldehyde, 0.5% glutaraldehyde in 0.1 M cacodylate buffer at pH 7.4, also contained 2 mM of CaCl2 , 1 mM of MgCl2, and 0.1% of tannic acid for conventional observation; and fuether 30-90 mM of potassium oxalate containing fixative was applied for the detection of calcium ion localization in cells. Tissue blocks were left in the same fixative for 30 to 180 min after MWI at room temperature, then proceeded to the sampling procedure, after postfixed with osmium tetroxide, embedded in Epon. Ultrathin sections were double stained with an useal manner. Oxalate treated sections were devided in two, stained and unstained one. The later oxalate treated unstained sections were analyzed with electron probe X-ray microanalyzer, the EDAX-PU-9800, at 40 KV accelerating voltage for 100 to 200 sec with point or selected area analyzing methods.

An ultrastructural study of sertoli cells in two geographically isolated populations of Aphanius dispar

1992 ◽  
Vol 50 (1) ◽  
pp. 548-549
Author(s):  
Taber A. Ba-Omar ◽  
Philip F. Prentis
Keyword(s):  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Freshwater Teleost ◽  
Isolated Populations ◽  
En Bloc ◽  
The North ◽  
Group A ◽  
Ultrathin Sections ◽  
Group B ◽  
Geographically Isolated

We have recently carried out a study of spermiogenic differentiation in two geographically isolated populations of Aphanius dispar (freshwater teleost), with a view to ascertaining variation at the ultrastructural level. The sampling areas were the Jebel Al Akhdar in the north (Group A) and the Dhofar region (Group B) in the south. Specimens from each group were collected, the testes removed, fixed in Karnovsky solution, post fixed in OsO, en bloc stained with uranyl acetate and then routinely processed to Agar 100 resin, semi and ultrathin sections were prepared for study.

The Identification of Phospholipid Micelles in Glutaraldehyde-Urea Embedded Tissue

1975 ◽  
Vol 33 ◽  
pp. 494-495
Author(s):  
Daniel C. Pease
Keyword(s):  
Electron Micrographs ◽  
Lung Tissue ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Type Ii ◽  
Lead Citrate ◽  
Phospholipid Micelles ◽  
Type Ii Pneumocytes ◽  
Ultrathin Sections ◽  
Polar Water

It is reasonable to think that phospholipid micelles should be visible and identifiable in electron micrographs of ultrathin sections if only they can be preserved throughout the embedding process. The development of highly polar, water-containing, aminoplastic embedments has made this a likely possibility. With this in mind, an investigation of the lecithin-secreting, Type II pneumocytes of the lung is underway.Initially it has been easiest to recognize phospholipid micelles in lung tissue fixed first with glutaraldehyde, and then secondarily exposed to osmium tetroxide. However, the latter is not a necessary concomitant for micellar preservation. Conventional uranyl acetate and lead citrate staining is finally applied. Importantly, though, the micelles have been most easily seen in tissue embedded in 507. glutaraldehyde polymerized with urea, as described in detail by D.C. Pease and R.G. Peterson (J. Ultra- struct. Res., 41, 133, 1972). When oriented appropriately, the micellar units are seen as tiny, bilayer plates.

Ultrastructural morphology of neurosecretory neurons in the barnacle Balanus amphitrite

1990 ◽  
Vol 48 (3) ◽  
pp. 434-435
Author(s):  
Grazia Tagliafierro ◽  
Cristiana Crosa ◽  
Marco Canepa ◽  
Tiziano Zanin
Keyword(s):  
Nervous System ◽  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Balanus Amphitrite ◽  
Neurosecretory Neurons ◽  
The Central Nervous System ◽  
Ultrathin Sections ◽  
Dense Core Granules ◽  
The Brain ◽  
Ocellar Nerve

Barnacles are very specialized Crustacea, with strongly reduced head and abdomen. Their nervous system is rather simple: the brain or supra-oesophageal ganglion (SG) is a small bilobed structure and the toracic ganglia are fused into a single ventral mass, the suboesophageal ganglion (VG). Neurosecretion was shown in barnacle nervous system by histochemical methods and numerous putative hormonal substances were extracted and tested. Recently six different types of dense-core granules were visualized in the median ocellar nerve of Balanus hameri and serotonin and FMRF-amide like substances were immunocytochemically detected in the nervous system of Balanus amphitrite. The aim of the present work is to localize and characterize at ultrastructural level, neurosecretory neuron cell bodies in the VG of Balanus amphitrite.Specimens of Balanus amphitrite were collected in the port of Genova. The central nervous system were Karnovsky fixed, osmium postfixed, ethanol dehydrated and Durcupan ACM embedded. Ultrathin sections were stained with uranyl acetate and lead citrate. Ultrastructural observations were made on a Philips M 202 and Zeiss 109 T electron microscopy.

Activation of bovine oocytes matured in vitro by injection of bovine and human spermatozoa or their cytosolic fractions

Zygote ◽  
2001 ◽  
Vol 9 (1) ◽  
pp. 89-95 ◽  
Author(s):  
Osamu Okitsu ◽  
Shuji Yamano ◽  
Toshihiro Aono
Keyword(s):  
Human Sperm ◽  
Human Spermatozoa ◽  
Oocyte Activation ◽  
Sham Injection ◽  
Bovine Sperm ◽  
Female Pronucleus ◽  
Sperm Factor ◽  
Bovine Spermatozoa ◽  
Bovine Oocytes

The aim of this study was to investigate whether bovine spermatozoa possess so-called sperm factor in the cytosolic fraction (CF) which activates bovine oocytes, and whether bovine oocytes matured in vitro are activated by microinjection of CF extracted from spermatozoa of other species. In the first experiment, bovine and human spermatozoa were microinjected into ooplasm of bovine oocytes matured in vitro. Secondly, CF from bovine and human spermatozoa were injected into bovine oocytes. In the third, CF from human spermatozoa was injected into human unfertilised oocytes obtained 18-20 h after clinical intracytoplasmic sperm injection (ICSI). We found that microinjection of bovine spermatozoa into bovine oocytes induced oocyte activation, as shown by resumption of meiosis and formation of a female pronucleus, at a significantly higher rate than the bovine sham injection (63.0% vs 43.0%; p < 0.05). On the other hand, there was no significant difference in activation rate between the human sperm injection (35.9%) and the human sham injection (22.9%). Furthermore, microinjection of bovine sperm CF into bovine oocytes induced oocyte activation at a significantly higher rate than the human CF injection or sham injection (75.9% vs 14.8%, 20.4%; p < 0.01). Formation of a single female pronucleus and second polar body extrusion was observed in 95.1% of activated oocytes after bovine sperm CF injection. When human sperm CF was injected into human unfertilised oocytes, the activation rate was significantly higher than following sham injection (76.9% vs 44.0%; p < 0.05). These results indicate the presence of sperm factor in bovine sperm CF which activate bovine oocytes, and suggest the possibility that sperm factor has species-specificity at least between bovine and human.

Ultrastructural evaluation of the effects induced by a macrolide antibiotic (LY281389) on L6 muscle cells in vitro

1991 ◽  
Vol 49 ◽  
pp. 80-81
Author(s):  
J. W. Horn ◽  
S. L. White ◽  
D. A. Laska ◽  
M. K. Buening ◽  
M. N. Novilla ◽  
...  
Keyword(s):  
Macrolide Antibiotic ◽  
Uniform Density ◽  
Solution Ph ◽  
L6 Cells ◽  
Vacuolar Change ◽  
Interference Range ◽  
Cacodylate Buffer ◽  
Ultrathin Sections ◽  
L6 Muscle Cells

Toxic doses of LY281389 (9-N(n-propyl)erythromycylamine), a cationic amphophilic macrolide antibiotic, induce a generalized cytoplasmic vacuolar change in tissues of rats and dogs. The present in vitro study using L6 cells (a line derived from neoplastic skeletal muscle) was done to evaluate sequentially early cytologic changes.The L6 cells seeded on tissue culture plates were allowed to grow to a uniform density. The culture medium was then replaced with control medium or medium containing 0.25 mg/ml LY281389 for exposures of 0.5, 2, 6, 12, 24, or 48 hours. The L6 cells were then fixed for 1-hour in modified Karnovsky's solution (pH 7.2), rinsed with 0.1 M sodium cacodylate buffer (pH 7.2) and postfixed for an additional hour in 2% osmium tetroxide (pH 7.2). Following a second buffer rinse, the L6 cells were dehydrated in graded ethanol solutions. The cells on each plate were embedded with epoxy resin (EPON 812), selected areas were scribed out and re-embedded for orientation. Ultrathin sections (silver to gold color interference range) were mounted on copper 200 mesh grids, counterstained, and examined using a Philips EM410LS electron microscope at 60kV.

Ultrastructural studies of cartilage in genetic disorders of skeletal growth

1978 ◽  
Vol 36 (2) ◽  
pp. 668-669
Author(s):  
D. O. Sillence ◽  
D. L. Rimoin ◽  
Ruth Silberberg
Keyword(s):  
Genetic Disorders ◽  
Electron Microscopic Examination ◽  
Uranyl Acetate ◽  
Osmic Acid ◽  
Electron Microscopic ◽  
Skeletal Dysplasias ◽  
Ultrastructural Studies ◽  
Low Viscosity ◽  
Transmission Electron ◽  
Cacodylate Buffer

The human skeletal dysplasias are an heterogeneous group of heritable connective tissue disorders associated with abnormalities in the size and shape of the limbs, trunk and/or skull which frequently result in disproportionate short stature. In recent years it has become apparent that these comprise over 50 distinct conditions with a variety of subtypes distinguished on clinical and radiological grounds. We have investigated the pathogenesis of these conditions in over 100 patients by direct transmission electron microscopic examination of chondro-osseous tissue. Some of the ultrastructural studies have been previously reported.Small biopsies of chondro-osseous junction were collected for electron microscopy from the rib or iliac crest of patients with skeletal dysplasias or from normal controls at the time of surgery. These were cut into small blocks and fixed for one hour in either 5% glutaraldehyde in white's buffer or directly in 1% osmic acid in White's buffer or a modified Karnovsky's fixative, (2. 5% paraformaldehyde, 2. 5% glutaraldehyde, 2. 5mM calcium in cacodylate buffer). Subsequent processing included osmium fixation, block staining with uranyl acetate and embedding in Araldite or Spurr's low viscosity resin (firm composition). Sections were cut with glass knives or diamond knives. The latter produced sections which were much more even in thickness, permitting more consistent appraisal of matrix features.

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