An improved method for ultrastructural examination of paraffin-embedded tissues for diagnostic purposes

1993 ◽  
Vol 51 ◽  
pp. 32-33
Author(s):  
Tamara A. Howard ◽  
Donna N. Ziesmer ◽  
Ardith L. Ries ◽  
Michael J. Becich
Keyword(s):  
Metabolic Disorders ◽  
Cell Membranes ◽  
Lipid Extraction ◽  
Soft Tissue Tumors ◽  
Ultrastructural Examination ◽  
Polar Solvents ◽  
Tissue Samples ◽  
Cacodylate Buffer ◽  
Skin Biopsies ◽  
Soluble Components

Small cell tumors, liver and skin biopsies for metabolic disorders, soft tissue tumors and kidney biopsies are the paraffin embedded tissues most frequently requested for ultrastructural examination. Preservation of cell membranes, glycogen, mucopolysaccharides, glycoproteins, glycolipids and other soluble components is problematic in these cases. Causes of cellular extraction when processing paraffin embedded tissue are:a) inadequate fixation of carbohydrates, mucopolysaccarides, glycoproteins and glycolipidsb) extraction of lipids and cell membranes by polar solventsc) temperature effectsd) extraction of sugars and glycoproteins due to pH and non-buffered, hypo-osmolar solutions.A methodology which improves preservation is described below:1) Tissue samples are cut from paraffin blocks and deparaffinized in 2 changes of xylene at 4°C for 2-4 h (low temperature decreases extraction in solvent).2) The samples are then rehydrated through a decreasing series of acetone concentrations (5-10 min each) at 4°C to 2, 5-10 min changes of 0.1 M cacodylate buffer containing 14.4 mM sucrose and 3.5 mM CaCl2, pH 7.8 (less polar solvent -- acetone vs. ethanol -- should decrease lipid extraction thereby enhancing membrane preservation)

Oligodendrocytes in Developing Hameter Cerebral Cortex

1976 ◽  
Vol 34 ◽  
pp. 126-127
Author(s):  
MB. Tank Buschmann
Keyword(s):  
Cerebral Cortex ◽  
Optic Nerve ◽  
Frontal Cortex ◽  
Osmium Tetroxide ◽  
Sodium Cacodylate Buffer ◽  
Sodium Cacodylate ◽  
Tissue Samples ◽  
Cacodylate Buffer ◽  
Layer V ◽  
Adult Hamster

Development of oligodendrocytes in rat corpus callosum was described as a sequential change in cytoplasmic density which progressed from light to medium to dark (1). In rat optic nerve, changes in cytoplasmic density were not observed, but significant changes in morphology occurred just prior to and during myelination (2). In our study, the ultrastructural development of oligodendrocytes was studied in newborn, 5-, 10-, 15-, 20-day and adult frontal cortex of the golden hamster (Mesocricetus auratus).Young and adult hamster brains were perfused with paraformaldehyde-glutaraldehyde in sodium cacodylate buffer at pH 7.3 according to the method of Peters (3). Tissue samples of layer V of the frontal cortex were post-fixed in 2% osmium tetroxide, dehydrated in acetone and embedded in Epon-Araldite resin.

Nasal Chondromesenchymal Hamartoma in Children

2001 ◽  
Vol 125 (3) ◽  
pp. 400-403 ◽  
Author(s):  
Chuen Hsueh ◽  
Swei Hsueh ◽  
Frank Gonzalez-Crussi ◽  
Ta-jen Lee ◽  
Jen-liang Su
Keyword(s):  
Immunohistochemical Study ◽  
Correct Diagnosis ◽  
S100 Protein ◽  
Soft Tissue Tumors ◽  
Biological Behavior ◽  
Ultrastructural Examination ◽  
Spindle Cells ◽  
Nasal Chondromesenchymal Hamartoma ◽  
Myofibroblastic Differentiation ◽  
Malignant Soft Tissue

Abstract Hamartoma in the nasal cavity of children is especially rare. Most documented cases occurred in infants, with characteristic histologic features of a mixture of various mesenchymal tissues. McDermott et al designated it nasal chondromesenchymal hamartoma in 1998, and it has since been considered a distinct clinicopathological entity. We report 2 such examples in a full-term male newborn and a 9-month-old boy, respectively. Histologically, both cases were characterized by a mixture of various mesenchymal elements, including spindle cells, collagen fibers, and irregular islands of osseous and chondroid tissue. Immunohistochemical study showed positivity to vimentin and S100 protein. Ultrastructural examination of case 1 demonstrated fibroblastic and myofibroblastic differentiation in tumor cells. There were 11 cases of nasal chondromesenchymal hamartoma in children published to date. The tumor has a benign biological behavior, and complete resection is the treatment of choice. It is apt to be misdiagnosed because of overlapping histologic features shared with a number of benign and malignant soft tissue tumors. Awareness of this entity is essential for correct diagnosis and adequate therapy.

scholarly journals An Untypical Case of Gouty Infiltration of Both Peroneal Tendons and a Longitudinal Lesion of the Peroneus Brevis Tendon Mimicking Synovial Sarcoma

2018 ◽  
Vol 2018 ◽  
pp. 1-3 ◽  
Author(s):  
Konstantinos Anagnostakos ◽  
Andreas Thiery ◽  
Christof Meyer ◽  
Octavian Tapos
Keyword(s):  
Synovial Sarcoma ◽  
Histopathological Examination ◽  
Soft Tissue Tumors ◽  
Resonance Imaging ◽  
Open Biopsy ◽  
Tissue Samples ◽  
Peroneal Tendons ◽  
Orthopedic Surgeons ◽  
History Of ◽  
Peroneus Brevis

We present a case of a 70-year-old male patient with an untypical gout infiltration of the peroneal tendons mimicking synovial sarcoma. The patient had a negative history of gout at initial presentation in our department. Magnetic resonance imaging of the region revealed a finding highly suspicious for synovial sarcoma of the peroneal tendons. Open biopsy was performed. Histopathological examination of the tissue samples demonstrated the presence of gout with no signs of malignancy. The gout infiltration was excised in a subsequent surgery. Orthopedic surgeons should be aware of the potential manifestation of gout in tendons and bear this in mind in the differential diagnosis of soft tissue tumors.

scholarly journals Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy

2011 ◽  
Vol 68 (5) ◽  
pp. 455-459 ◽  
Author(s):  
Zeljko Krsmanovic ◽  
Evica Dincic ◽  
Smiljana Kostic ◽  
Vesna Lackovic ◽  
Milos Bajcetic ◽  
...  
Keyword(s):  
Autosomal Dominant ◽  
Case Reports ◽  
Pathological Process ◽  
Unknown Etiology ◽  
Cognitive Deterioration ◽  
Distinctive Pattern ◽  
Ultrastructural Examination ◽  
Notch 3 ◽  
Skin Biopsies ◽  
The Brain

Introduction. Fast and precise diagnostics of the disease from the large group of adult leukoencephalopathy is difficult but responsible job, because the outcome of the disease is very often determined by its name. Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is caused by the mutation of Notch 3 gene on chromosome locus 19p13. Beside the brain arterioles being the main disease targets, extracerebral small blood vessels are affected by the pathological process. Clinically present signs are recurrent ischemic strokes and vascular dementia. CADASIL in its progressive form shows a distinctive pattern of pathological changes on MRI of endocranium. The diagnosis is confirmed by the presence of granular osmiophilic material (GOM) in histopathological skin biopsies. Case reports. Two young adult patients manifested ischemic strokes of unknown etiology, cognitive deterioration, migraine and psychopathological phenomenology. MRI of endocranium pointed on CADASIL. Ultrastructural examination of skin biopsy proved the presence of GOM in the basal lamina and near smooth muscle cells of arteriole dermis leading to CADASIL diagnosis. The presence of GOM in histopathological preparation is 100% specific for CADASIL. The patients were not searched for mutation in Notch 3 gene on chromosome 19, because some other leukoencephalopathy was disregarded. Conclusion. Suggestive clinical picture, distinctive finding of endocranium MRI, the presence of GOM by ultrastructural examination of histopathological skin biopsies are sufficient to confirm CADASIL diagnosis.

Development of iron granules in honey bee oenocytes

1984 ◽  
Vol 42 ◽  
pp. 744-745
Author(s):  
Deborah A. Kuterbach
Keyword(s):  
Honey Bee ◽  
Honey Bees ◽  
Osmium Tetroxide ◽  
Developmental Stages ◽  
Iron Particles ◽  
Sodium Cacodylate ◽  
Ultrastructural Examination ◽  
Brood Comb ◽  
Cacodylate Buffer ◽  
Rich Material

Foraging honey bees are believed to use the earth's magnetic field, among other cues, in order to home. It has been reported that the abdomen of the honey bee contains magnetite and iron particles have been localized within abdominal oenocytes. Light microscopic investigations reveal that morphologically detectable iron granules are present only in adult animals older than six days after eclosion (emergence from the comb). This is a report of an ultrastructural examination of the oenocytes during the development of the worker honey bee (Apis mellifera) with particular emphasis on the time of appearance, number, and size of iron granules within the cells.Specimens of the different developmental stages were removed from brood comb, fixed in 2.5% glutaraldehyde in 5mM sodium cacodylate buffer pH 7.3, washed, and post-fixed with 1% osmium tetroxide. In order to preserve the lipid-rich material, rapid dehydration was accomplished by three changes of 50% acetone and two changes of 100% acetone before embedding in Polybed 812 epoxy resin.

Electron Diffraction Studies in Mercury Neuropoisoning

1976 ◽  
Vol 34 ◽  
pp. 270-271
Author(s):  
W. W. Shelton ◽  
T. K. Chatterjee
Keyword(s):  
Body Weight ◽  
Electron Diffraction ◽  
Uranyl Acetate ◽  
Granule Cell Layer ◽  
Sprague Dawley ◽  
Tissue Samples ◽  
Subcutaneous Injections ◽  
Sprague Dawley Rats ◽  
Cacodylate Buffer ◽  
Cerebellar Tissue

Mercurials have been shown to accumulate in the vertebrate cerebellum and ultimately induce cytological pathology, especially in the granule cell layer, under conditions of experimental poisoning (1). This report describes some preliminary results of electron diffraction and electron microscopy (Philips EM300) studies on cerebellar tissue of rats poisoned with mercuric bichloride (MB) (Fisher Scientific Co.) and phenylmercuric acetate (PMA) (Eastman). One group of male albino Sprague Dawley rats was treated with single subcutaneous injections of 10 mg. MB per kg. body weight and observed for one week, and another group intraperitoneally with 2 mg. PMA per kg. body weight three times a week for two months. The animals were anesthetized and perfused intracardially with 3% glutaraldehyde in 0.1M cacodylate buffer and the selected tissue samples were further processed according to standard methods, terminating with uranyl acetate and lead citrate staining.

Effect of Water on Mechanical Properties of Mineralized Tissue Composites

2006 ◽  
Vol 975 ◽  
Author(s):  
Amanpreet Kaur Bembey ◽  
Michelle L Oyen ◽  
Virginia L. Ferguson ◽  
Andrew J. Bushby ◽  
Alan Boyde
Keyword(s):  
Mechanical Properties ◽  
Soft Tissue ◽  
Mechanical Parameters ◽  
Mineralized Tissue ◽  
Mineral Phase ◽  
Polar Solvents ◽  
Tissue Samples ◽  
Mineralized Tissues ◽  
Tissue Shrinkage ◽  
Tissue Phase

ABSTRACTIn the current study, the effects of polar solvents on tissue volume and mechanical properties are considered. Area shrinkage measurements are conducted for mineralized, bone tissue samples soaked in polar solvents. Area shrinkage is used to calculate approximate linear and volume shrinkage. Results are compared with viscoelastic mechanical parameters for bone in the same solvents (as measured previously) and with both shrinkage measurements and mechanical data for nonmineralized tissues, as taken from the existing literature. As expected, the shrinkage of mineralized tissues is minimal when compared with shrinkage of nonmineralized tissues immersed in the same polar solvents. The mechanical changes in bone are also substantially less than in nonmineralized tissues. The largest stiffness values are found in shrunken bone samples (immersed in acetone and ethanol). The mineral phase in bone thus resists tissue shrinkage that would otherwise occur in the pure soft tissue phase.

Scanning Electron Microscopy of ischemic intestinal mucosa

1987 ◽  
Vol 45 ◽  
pp. 732-733
Author(s):  
Ron W. Millard ◽  
Y. Nakajima ◽  
M. Ashraf
Keyword(s):  
Intestinal Mucosa ◽  
Arterial Occlusion ◽  
Small Intestinal Mucosa ◽  
Cellular Viability ◽  
Tissue Samples ◽  
Cacodylate Buffer ◽  
Anesthetized Dogs ◽  
Small Intestinal ◽  
Scanning Electron ◽  
Made In

The small intestinal mucosa is known to be sensitive to ischemia. The early changes which are critical to the eventual cellular viability upon restoration of blood flow or other therapeutic intervention have not been investigated thoroughly. This investigation describes the surface alterations of the villous cells in the dog's small intestine. Jejunal segments of 14 anesthetized dogs were subjected to the arterial occlusion for 5, 15, 30, 45, and 60 minutes. At the end of the occlusion period, tissue samples from normal non-ischemic and ischemic (I) zones were fixed in 2.5% glutaraldehyde made in 0.17 M cacodylate buffer, pH 7.4, for four hours, rinsed in the buffer, and dehydrated in a graded series of ethanol. Finally, they were critically point-dried and coated with gold-palladium (60:40) for examination in the ETEC scanning electron microscope.

Membrane Organelles in Developing Hamster Frontal Cortex

1975 ◽  
Vol 33 ◽  
pp. 316-317
Author(s):  
MB. Tank Buschraann
Keyword(s):  
Frontal Cortex ◽  
Osmium Tetroxide ◽  
Lamellar Body ◽  
Pyramidal Cells ◽  
Tissue Samples ◽  
Cacodylate Buffer ◽  
Spine Apparatus ◽  
Subsurface Cisterns ◽  
Subsurface Cistern ◽  
Nissl Substance

Changes in the morphology of the protein-synthetic organelles in pyramidal cells from the frontal cortex were studied in the newborn, 5-, 10-, 15-, 20- day, and adult golden hamster (Mesocricetus auratus). The rough endoplasmic reticulum (RER) was of special interest since it forms the characteristic Nissl substance of adult neurons, as well as other occasional specializations including the subsurface cistern, the lamellar body, the nebenkem, the multilaminated body, the annulate lamella, and the spine apparatus. During the course of this investigation, a sequential pattern was noted in the appearance of the subsurface cisterns and the lamellar bodies, which were the only two specializations of RER observed.The animals were perfused with glutaraldehyde-paraformaldehyde in cacodylate buffer (pH 7.3) after the method of Peters. Tissue samples were subsequently post-fixed in 2% osmium tetroxide, dehydrated in acetone, and embedded in Epon-Araldite resin.

scholarly journals Lipid yield from the diatom Porosira glacialis is determined by solvent choice and number of extractions, independent of cell disruption

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Jon Brage Svenning ◽  
Lars Dalheim ◽  
Terje Vasskog ◽  
Lucie Matricon ◽  
Birthe Vang ◽  
...  
Keyword(s):  
Lipid Class ◽  
Lipid Extraction ◽  
Solvent Polarity ◽  
Polar Solvents ◽  
Lipid Class Composition ◽  
Lipid Yield ◽  
Class Composition ◽  
Cell Wall Disruption ◽  
Odontella Aurita ◽  
Main Factors

AbstractCell wall disruption is necessary to maximize lipid extraction yields in conventional species of mass-cultivated microalgae. This study investigated the effect of sonication, solvent choice and number of extractions on the lipid yield, lipid class composition and fatty acid composition of the diatom Porosira glacialis. For comparison, the diatom Odontella aurita and green alga Chlorella vulgaris were included in the study. Sonication effectively disrupted P. glacialis cells, but did not increase the total lipid yield compared to physical stirring (mixing). In all three microalgae, the content of membrane-associated glyco- and phosopholipids in the extracted lipids was strongly dependent on the solvent polarity. A second extraction resulted in higher yields from the microalgae only when polar solvents were used. In conclusion, choice of solvent and number of extractions were the main factors that determined lipid yield and lipid class composition in P. glacialis.

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