Effect of Acetone and Kerosene on Skin Ultrastructure

Author(s):  
A. P. Lupulescu ◽  
H. Pinkus ◽  
D. J. Birmingham

Our laboratory is engaged in the study of the effect of different chemical agents on human skin, using electron microscopy. Previous investigations revealed that topical use of a strong alkali (NaOH 1N) or acid (HCl 1N), induces ultrastructural changes in the upper layers of human epidermis. In the current experiments, acetone and kerosene, which are primarily lipid solvents, were topically used on the volar surface of the forearm of Caucasian and Negro volunteers. Skin specimens were bioptically removed after 90 min. exposure and 72. hours later, fixed in 3% buffered glutaraldehyde, postfixed in 1% phosphate osmium tetroxide, then flat embedded in Epon.

Author(s):  
Robert Giaquinta ◽  
M. A. Hayat

The ultrastructural changes that occur in the intestinal absorptive cells during amphibian metamorphosis have been reported (Bonneville, 1963). These changes accompany a change in diet (from an herbivorous to a carnivorous state) during metamorphosis. Little information is available, however, on the ultrastructural changes in the absorptive cells of amphibians in relation to the state of feeding. This report describes the differences in the fine structure of these cells in the tadpole stage of Rana pipiens during periods of food absorption and fasting.Rana pipiens at tadpole stages were fed an herbivorous diet, and after a period of 48 hr, the animal was dissected and segments of the small intestine were collected for electron microscopy. A second group of tadpoles was fasted for 7 days, and segments of the small intestine were collected. The tissue specimens were immersed in phosphate-buffered glutaraldehyde (3%) for 1 hr at 4C and postfixed with phosphate-buffered osmium tetroxide (2%) for 1 hr at 4C.


Author(s):  
D.L. Friesen ◽  
A. Singh ◽  
M.E. Hitt

Thiacetarsamide is an arsenic-containing drug used in the treatment of heartworm in dogs. The effective antihelmintic dose is toxic to the host animal. Acetylcysteine decreases the hepatotoxicity of some compounds by forming a conjugate with toxic metabolites of the compound. The purpose of this study was to evaluate the effectiveness of a cytoprotectant for hepatocytes in dogs treated with therapeutic levels of thiacetarsamide.Eighteen dogs were divided randomly into two groups. All dogs were given four doses of thiacetarsamide over two days. Nine dogs were given 10% acetylcysteine 15 min prior to each dose of thiacetarsamide. Needle biopsies of the liver were taken from each dog prior to the treatment and again one week post-treatment. The biopsies were fixed in 2% gluraraldehyde in phosphate buffer, pH 7.3, post-fixed in 1% osmium tetroxide, and processed for electron microscopy. Semithin and thin sections of the liver were examined by light and electron microscopy, respectively, for histopathologic and ultrastructural changes. The specimens were coded and the sample treatment was not known to the researchers at the time of observation.


1958 ◽  
Vol 4 (6) ◽  
pp. 679-684 ◽  
Author(s):  
Wallace H. Clark ◽  
Richard G. Hibbs

The human epidermis has been studied by electron microscopy following osmium tetroxide and potassium permanganate fixation. An anatomically distinct cell in the human epidermis has been demonstrated with features similar to the melanocyte of the hair bulb described by Barnicot, Birbeck and Cuckow (3). It is dendritic in form and does not contain tonofilaments. "Intercellular bridges" are not formed. The mitochondria are larger and more numerous than those of other epidermal cells and the endoplasmic reticulum is more complex. Some of these cells contain melanin but others are melanin-free. The cell has been interpreted as being identical with the dopa-positive, clear cell of Masson (dendritic cell of Bloch or melanocyte). We have found that many membranous structures in the human epidermis are better preserved by permanganate fixation than by osmium tetroxide fixation.


2013 ◽  
Vol 12 (3) ◽  
pp. 76-81
Author(s):  
T. V. Tupitsyna ◽  
S. V. Logvinov ◽  
O. A. Tikhonovskaya ◽  
M. L. Dmitrieva

Autoimmune oophoritis (AO) is characterized by damage of generative and endocrine elements of the ovaries and leads to the formation of secondary insufficiency of the gonads. A question about the use of glucocorticoids (GC) for the correction of AO remains debatable.Objective: to study the electron-microscopic changes of structural-tissue cells in autoimmune ovariano ophoritis and after its correction by glucocorticoids in the experiment.The material. The experiment was performed on outbred white Mature rats-female. The main group of animals (12 rats) were simulated AO by intra-peritoneal introduction of antigens of ovarian. At 5th day prednisolone was injected to rats («Nycomed», Austria) in the dose of 3 mg/kg of body weight intramuscularly 14-day course. A comparison group (12 rats) were the animals with the model AO no course Ledger therapy. Controlintact rats (6 animals). Taking of the material was carried out by the 20th and the 60th day.Methods. The study of the ultrastructure ovaries were performed using transmission electron microscopy. The material was fixed 2.5% glutaraldehyde, postfixed in 2% solution of osmium tetroxide, dehydrated in ethyl alcohols rising concentration and placed in a mixture of resins Epon-Araldite. Sections were prepared on Ultrotom III (LKB, Sweden). To view the drugs used an electron microscope JEM-7A (Japan). Results. On the 20th day of the pilot AO there are significant ultrastructural changes of vessels, concerning mainly endothelial layer. Degenerative and destructive changes affected endocrinocytes in the composition of the libraries and granular, овоциты majority of antral follicles. The 60-day pathologic processes involved and preantral follicles in the medulla develop perivascular fibrosis-sclerotic changes. Holding GC therapy on the 5th day AO reduces ultrastructural breach the walls of blood vessels, limits migration of immune cells in the home defeat on the 20th day. By the 60th day in the conditions of the restored the blood tissue transport is the formation of full-fledged generative elements, confirmed by the results of electron microscopy study.Conclusion. The obtained experimental data are demonstrated the therapeutic efficacy of the GC-therapy in the early stages of AO.


Author(s):  
Ronald H. Bradley ◽  
R. S. Berk ◽  
L. D. Hazlett

The nude mouse is a hairless mutant (homozygous for the mutation nude, nu/nu), which is born lacking a thymus and possesses a severe defect in cellular immunity. Spontaneous unilateral cataractous lesions were noted (during ocular examination using a stereomicroscope at 40X) in 14 of a series of 60 animals (20%). This transmission and scanning microscopic study characterizes the morphology of this cataract and contrasts these data with normal nude mouse lens.All animals were sacrificed by an ether overdose. Eyes were enucleated and immersed in a mixed fixative (1% osmium tetroxide and 6% glutaraldehyde in Sorenson's phosphate buffer pH 7.4 at 0-4°C) for 3 hours, dehydrated in graded ethanols and embedded in Epon-Araldite for transmission microscopy. Specimens for scanning electron microscopy were fixed similarly, dehydrated in graded ethanols, then to graded changes of Freon 113 and ethanol to 100% Freon 113 and critically point dried in a Bomar critical point dryer using Freon 13 as the transition fluid.


Author(s):  
Jane A. Westfall ◽  
S. Yamataka ◽  
Paul D. Enos

Scanning electron microscopy (SEM) provides three dimensional details of external surface structures and supplements ultrastructural information provided by transmission electron microscopy (TEM). Animals composed of watery jellylike tissues such as hydras and other coelenterates have not been considered suitable for SEM studies because of the difficulty in preserving such organisms in a normal state. This study demonstrates 1) the successful use of SEM on such tissue, and 2) the unique arrangement of batteries of nematocysts within large epitheliomuscular cells on tentacles of Hydra littoralis.Whole specimens of Hydra were prepared for SEM (Figs. 1 and 2) by the fix, freeze-dry, coat technique of Small and Màrszalek. The specimens were fixed in osmium tetroxide and mercuric chloride, freeze-dried in vacuo on a prechilled 1 Kg brass block, and coated with gold-palladium. Tissues for TEM (Figs. 3 and 4) were fixed in glutaraldehyde followed by osmium tetroxide. Scanning micrographs were taken on a Cambridge Stereoscan Mark II A microscope at 10 KV and transmission micrographs were taken on an RCA EMU 3G microscope (Fig. 3) or on a Hitachi HU 11B microscope (Fig. 4).


Author(s):  
M. O. Magnusson ◽  
D. G. Osborne ◽  
T. Shimoji ◽  
W. S. Kiser ◽  
W. A. Hawk

Short term experimental and clinical preservation of kidneys is presently best accomplished by hypothermic continuous pulsatile perfusion with cryoprecipitated and millipore filtered plasma. This study was undertaken to observe ultrastructural changes occurring during 24-hour preservation using the above mentioned method.A kidney was removed through a midline incision from healthy mongrel dogs under pentobarbital anesthesia. The kidneys were flushed immediately after removal with chilled electrolyte solution and placed on a LI-400 preservation system and perfused at 8-10°C. Serial kidney biopsies were obtained at 0-½-1-2-4-8-16 and 24 hours of preservation. All biopsies were prepared for electron microscopy. At the end of the preservation period the kidneys were autografted.


Author(s):  
J. G. Robertson ◽  
D. F. Parsons

The extraction of lipids from tissues during fixation and embedding for electron microscopy is widely recognized as a source of possible artifact, especially at the membrane level of cell organization. Lipid extraction is also a major disadvantage in electron microscope autoradiography of radioactive lipids, as in studies of the uptake of radioactive fatty acids by intestinal slices. Retention of lipids by fixation with osmium tetroxide is generally limited to glycolipids, phospholipids and highly unsaturated neutral lipids. Saturated neutral lipids and sterols tend to be easily extracted by organic dehydrating reagents prior to embedding. Retention of the more saturated lipids in embedded tissue might be achieved by developing new cross-linking reagents, by the use of highly water soluble embedding materials or by working at very low temperatures.


Author(s):  
Douglas R. Keene ◽  
Robert W. Glanville ◽  
Eva Engvall

A mouse monoclonal antibody (5C6) prepared against human type VI collagen (1) has been used in this study to immunolocalize type VI collagen in human skin. The enbloc method used involves exposing whole tissue pieces to primary antibody and 5 nm gold conjugated secondary antibody before fixation, and has been described in detail elsewhere (2).Biopsies were taken from individuals ranging in age from neonate to 65 years old. By immuno-electron microscopy, type VI collagen is found to be distributed as a fine branching network closely associated with (but not attached to) banded collagen fibrils containing types I and III collagen (Fig. 1). It appears to enwrap fibers, to weave between individual fibrils within a fiber, and to span the distance separating fibers, creating a “web-like network” which entraps fibers within deep papillary and reticular dermal layers (Fig. 2). Relative to that in the dermal matrix, the concentration of type VI collagen is higher around endothelial basement membranes limiting the outer boundaries of nerves, capillaries, and fat cells (Fig. 3).


Author(s):  
John H. Luft

With information processing devices such as radio telescopes, microscopes or hi-fi systems, the quality of the output often is limited by distortion or noise introduced at the input stage of the device. This analogy can be extended usefully to specimen preparation for the electron microscope; fixation, which initiates the processing sequence, is the single most important step and, unfortunately, is the least well understood. Although there is an abundance of fixation mixtures recommended in the light microscopy literature, osmium tetroxide and glutaraldehyde are favored for electron microscopy. These fixatives react vigorously with proteins at the molecular level. There is clear evidence for the cross-linking of proteins both by osmium tetroxide and glutaraldehyde and cross-linking may be a necessary if not sufficient condition to define fixatives as a class.


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