Improved preservation of the otolithic membranes

1987 ◽  
Vol 45 ◽  
pp. 914-915
Author(s):  
G. M. Cohen ◽  
J. S. Grasso ◽  
M. L. Domeier ◽  
P. T. Mangonon
Keyword(s):  
Toluidine Blue ◽  
Potassium Phosphate ◽  
Vestibular Function ◽  
Spatial Relations ◽  
White Leghorn ◽  
Supporting Cells ◽  
Semithin Sections ◽  
Micromechanical Models ◽  
Soluble Components

Any explanation of vestibular micromechanics must include the roles of the otolithic and cupular membranes. However, micromechanical models of vestibular function have been hampered by unresolved questions about the microarchitectures of these membranes and their connections to stereocilia and supporting cells. Otolithic membranes are notoriously difficult to preserve because of severe shrinkage and loss of soluble components. We have empirically developed fixation procedures that reduce shrinkage artifacts and more accurately depict the spatial relations between the otolithic membranes and the ciliary bundles and supporting cells.We used White Leghorn chicks, ranging in age from newly hatched to one week. The inner ears were fixed for 3-24 h in 1.5-1.75% glutaraldehyde in 150 mM KCl, buffered with potassium phosphate, pH 7.3; when postfixed, it was for 30 min in 1% OsO4 alone or mixed with 1% K4Fe(CN)6. The otolithic organs (saccule, utricle, lagenar macula) were embedded in Araldite 502. Semithin sections (1 μ) were stained with toluidine blue.

Fine structure of the cupula

1987 ◽  
Vol 45 ◽  
pp. 826-827
Author(s):  
G. M. Cohen ◽  
M. L. Domeier
Keyword(s):  
Fine Structure ◽  
Hair Cell ◽  
Dynamic Range ◽  
Potassium Phosphate ◽  
Basic Solution ◽  
White Leghorn ◽  
Original Volume ◽  
Old Mice ◽  
Stimulation Of

The cupula functions as a sensitive biological transducer that undergoes extremely limited mechanical displacements in its normal dynamic range. Cupular displacements are coupled to the bending and stimulation of hair cell cilia. However, details of cupular-ciliary coupling and of cupular attachments to the ampullary crest are unsettled because of difficulties in preserving the cupula without severe distortion from fixation and dehydration. With conventional fixation procedures, the cupula either pulls away from crest or collapses to a fraction of its original volume. Our objective was to reduce cupular shrinkage.We used the inner ears of 1 day-old chicks (White Leghorn) and 3 month-old mice (C57BL/6). The inner ears were fixed 3-72 h in 1.5-1.75% glutaraldehyde in 150 mM KC1 buffered with potassium phosphate, pH 7.3. To the basic solution, we sometimes added spermine (0.1%) or lysine (0.25%).

Sclerotome Mitotic Activity in Stage 19 Chick Embryo Braquial Somites

1997 ◽  
Vol 3 (S2) ◽  
pp. 179-180
Author(s):  
M. González-Santander Martinez ◽  
M. Monteagudo de la Rosa ◽  
G. Martinez Cuadrado ◽  
R. González Santander
Keyword(s):  
Extracellular Matrix ◽  
Cell Differentiation ◽  
Chick Embryo ◽  
Mitotic Activity ◽  
Toluidine Blue ◽  
Light Microscope ◽  
Osmium Tetroxide ◽  
Chick Embryos ◽  
White Leghorn ◽  
Intercellular Spaces

Somite disintegration begins with the disruption and dispersion of cells in its ventromedial region, that upon detaching from the somite form a defined group of cells called “sclerotome”. The sclerotome is formed by scleroblasts that originate paraxial segments. Scleroblasts disperse and migrate forming the “sclerotomal mesenchyma”. This sclerotomal mesenchyma, in its evolution and cell differentiation, will give rise to skeletal tissues. One of the first features of the scleroblast is the production of extracellular matrix rich in hialuronic acid that facilitates the spreading of filopodia and the migration of the sclerotome. Sclerotome development is closely associated to the microenvironment created by the extracellular matrix in which scleroblasts are located. Hialuronic acid is found in the ample intercellular spaces occupied by the hydrated matrix.White Leghorn chick embryos at H.H. stages 9 to 12 were fixed in 2.5% glutaraldehyde, postfixed in 1 % osmium tetroxide and embedded in araldite. Semithin transversal sections were stained with toluidine blue and visualized under the light microscope to locate the sclerotome.

scholarly journals Regulation of Cellular Calcium in Vestibular Supporting Cells by Otopetrin 1

2010 ◽  
Vol 104 (6) ◽  
pp. 3439-3450 ◽  
Author(s):  
Euysoo Kim ◽  
Krzysztof L. Hyrc ◽  
Judith Speck ◽  
Yunxia W. Lundberg ◽  
Felipe T. Salles ◽  
...  
Keyword(s):  
Culture System ◽  
Calcium Concentration ◽  
Vestibular Function ◽  
Cytosolic Calcium ◽  
Extracellular Calcium ◽  
Dependent Manner ◽  
Supporting Cells ◽  
Calcium Dependent ◽  
Organ Culture System ◽  
High Concentrations

Otopetrin 1 (OTOP1) is a multitransmembrane domain protein, which is essential for mineralization of otoconia, the calcium carbonate biominerals required for vestibular function, and the normal sensation of gravity. The mechanism driving mineralization of otoconia is poorly understood, but it has been proposed that supporting cells and a mechanism to maintain high concentrations of calcium are critical. Using Otop1 knockout mice and a utricular epithelial organ culture system, we show that OTOP1 is expressed at the apex of supporting cells and functions to increase cytosolic calcium in response to purinergic agonists, such as adenosine 5′-triphosphate (ATP). This is achieved by blocking mobilization of calcium from intracellular stores in an extracellular calcium-dependent manner and by mediating influx of extracellular calcium. These data support a model in which OTOP1 acts as a sensor of the extracellular calcium concentration near supporting cells and responds to ATP in the endolymph to increase intracellular calcium levels during otoconia mineralization.

A Simple Dichromatic Stain for Plastic Embedded Tissues

1970 ◽  
Vol 28 ◽  
pp. 296-297 ◽  
Author(s):  
G. R. Mackay ◽  
M. L. Mead
Keyword(s):  
Electron Microscopy ◽  
Toluidine Blue ◽  
Biological Material ◽  
Good Reproducibility ◽  
Routine Work ◽  
Staining Techniques

Color contrasting of 1 to 2 micron sections of plastic embedded biological material is an important adjunct to electron microscopy. The procedures in general use today are simple and rapid giving monochromatic results, e.g., toluidine blue. Although many di- and polychromatic histologic staining techniques have been modified to obtain a counterstaining effect with plasticembedded tissue, the methods are usually undesirable for routine work because they are time consuming, complicated and often defy good reproducibility.

The Alteration of the Pore Spaces in Sandstones

1973 ◽  
Vol 31 ◽  
pp. 210-211
Author(s):  
R. E. Ferrell ◽  
G. G. Paulson
Keyword(s):  
Electron Microscope ◽  
Scanning Electron Microscope ◽  
The Other ◽  
Coarse Grained ◽  
Depositional Fabric ◽  
Soluble Components ◽  
Scanning Electron ◽  
Shape And Size

The pore spaces in sandstones are the result of the original depositional fabric and the degree of post-depositional alteration that the rock has experienced. The largest pore volumes are present in coarse-grained, well-sorted materials with high sphericity. The chief mechanisms which alter the shape and size of the pores are precipitation of cementing agents and the dissolution of soluble components. Each process may operate alone or in combination with the other, or there may be several generations of cementation and solution.The scanning electron microscope has ‘been used in this study to reveal the morphology of the pore spaces in a variety of moderate porosity, orthoquartzites.

Ultrastructure of Melanin Formation in Curvularia sp., Alternaria sp., and Drechslera Sorokiniana

1977 ◽  
Vol 35 ◽  
pp. 398-399
Author(s):  
M. H. Wheeler ◽  
W. J. Tolmsoff ◽  
A. A. Bell
Keyword(s):  
Potassium Phosphate ◽  
Thielaviopsis Basicola ◽  
Wild Type ◽  
Potassium Phosphate Buffer ◽  
Transmission Microscopy ◽  
Electron Transmission ◽  
Melanin Formation ◽  
Before And After ◽  
Alternaria Sp ◽  
Electron Transmission Microscopy

(+)-Scytalone [3,4-dihydro-3,6,8-trihydroxy-l-(2Hj-naphthalenone] and 1,8-di- hydroxynaphthalene (DHN) have been proposed as intermediates of melanin synthesis in the fungi Verticillium dahliae (1, 2, 3, 4) and Thielaviopsis basicola (4, 5). Scytalone is enzymatically dehydrated by V. dahliae to 1,3,8-trihydroxynaphthalene which is then reduced to (-)-vermelone [(-)-3,4- dihydro-3,8-dihydroxy-1(2H)-naphthalenone]. Vermelone is subsequently dehydrated to DHN which is enzymatically polymerized to melanin.Melanin formation in Curvularia sp., Alternaria sp., and Drechslera soro- kiniana was examined by light and electron-transmission microscopy. Wild-type isolates of each fungus were compared with albino mutants before and after treatment with 1 mM scytalone or 0.1 mM DHN in 50 mM potassium phosphate buffer, pH 7.0. Both chemicals were converted to dark pigments in the walls of hyphae and conidia of the albino mutants. The darkened cells were similar in appearance to corresponding cells of the wild types under the light microscope.

Dexamethasone Effect on Embryonic Chick Respiratory Epithelium

1982 ◽  
Vol 40 ◽  
pp. 248-249
Author(s):  
M.R. Richter ◽  
R.V. Blystone
Keyword(s):  
Lung Development ◽  
Critical Role ◽  
Respiratory Epithelium ◽  
Chick Embryos ◽  
Embryonic Chick ◽  
White Leghorn ◽  
Lung Maturation ◽  
Synthetic Analogs ◽  
Lung Physiology ◽  
Avian Lung

Dexamethasone and other synthetic analogs of corticosteroids have been employed clinically as enhancers of lung development. The mechanism(s) by which this steroid induction of later lung maturation operates is not clear. This study reports the effect on lung epithelia of dexamethasone administered at different intervals during development. White Leghorn chick embryos were used so as to remove possible maternal and placental influences on the exogenously applied steroid. Avian lung architecture does vary from mammals; however, respiratory surfactant produced by the lung epithelia serves an equally critical role in avian lung physiology.

High resolution surface structure of foot-and-mouth disease virus

1978 ◽  
Vol 36 (2) ◽  
pp. 376-377
Author(s):  
S. S. Breese ◽  
H. L. Bachrach
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Surface Structure ◽  
Disease Virus ◽  
Potassium Phosphate ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Physical Measurements ◽  
Mouth Disease Virus ◽  
Foot And Mouth

Models for the structure of foot-and-mouth disease virus (FMDV) have been proposed from chemical and physical measurements (Brown, et al., 1970; Talbot and Brown, 1972; Strohmaier and Adam, 1976) and from rotational image-enhancement electron microscopy (Breese, et al., 1965). In this report we examine the surface structure of FMDV particles by high resolution electron microscopy and compare it with that of particles in which the outermost capsid protein VP3 (ca. 30, 000 daltons) has been split into smaller segments, two of which VP3a and VP3b have molecular weights of about 15, 000 daltons (Bachrach, et al., 1975).Highly purified and concentrated type A12, strain 119 FMDV (5 mg/ml) was prepared as previously described (Bachrach, et al., 1964) and stored at 4°C in 0. 2 M KC1-0. 5 M potassium phosphate buffer at pH 7. 5. For electron microscopy, 1. 0 ml samples of purified virus and trypsin-treated virus were dialyzed at 4°C against 0. 2 M NH4OAC at pH 7. 3, deposited onto carbonized formvar-coated copper screens and stained with phosphotungstic acid, pH 7. 3.

A SEM and Light Microscope Study of the Epidermal Leaf Structures of the Carnivorous Plant, Sarracenia purpurea, L.

1971 ◽  
Vol 29 ◽  
pp. 358-359
Author(s):  
B. J. Panessa ◽  
J. F. Gennaro
Keyword(s):  
Methylene Blue ◽  
Toluidine Blue ◽  
Outer Surface ◽  
Microscope Study ◽  
Light Microscope ◽  
Carnivorous Plant ◽  
Sarracenia Purpurea ◽  
Inner Surface

Tissue from the hood and sarcophagus regions were fixed in 6% glutaraldehyde in 1 M.cacodylate buffer and washed in buffer. Tissue for SEM was partially dried, attached to aluminium targets with silver conducting paint, carbon-gold coated(100-500Å), and examined in a Kent Cambridge Stereoscan S4. Tissue for the light microscope was post fixed in 1% aqueous OsO4, dehydrated in acetone (4°C), embedded in Epon 812 and sectioned at ½u on a Sorvall MT 2 ultramicrotome. Cross and longitudinal sections were cut and stained with PAS, 0.5% toluidine blue and 1% azure II-methylene blue. Measurements were made from both SEM and Light micrographs.The tissue had two structurally distinct surfaces, an outer surface with small (225-500 µ) pubescent hairs (12/mm2), numerous stoma (77/mm2), and nectar glands(8/mm2); and an inner surface with large (784-1000 µ)stiff hairs(4/mm2), fewer stoma (46/mm2) and larger, more complex glands(16/mm2), presumably of a digestive nature.

Immunocytochemical detection of hepatic carbohydrate metabolic enzymes: Improved resolution with polyethylene glycol embedding and visio-bond semithin sections

1992 ◽  
Vol 50 (1) ◽  
pp. 792-793
Author(s):  
Kuixiong Gao ◽  
Randal E. Morris ◽  
Bruce F. Giffin ◽  
Robert R. Cardell
Keyword(s):  
Polyethylene Glycol ◽  
Glycogen Phosphorylase ◽  
Phosphoenolpyruvate Carboxykinase ◽  
Glycogen Synthase ◽  
Metabolic Enzymes ◽  
Silver Stain ◽  
Accurate Localization ◽  
Immunocytochemical Detection ◽  
Semithin Sections ◽  
Parenchymal Cells

Several enzymes are involved in the regulation of anabolic and catabolic pathways of carbohydrate metabolism in liver parenchymal cells. The lobular distribution of glycogen synthase (GS), phosphoenolpyruvate carboxykinase (PEPCK) and glycogen phosphorylase (GP) was studied by immunocytochemistry using cryosections of normal fed and fasted rat liver. Since sections of tissue embedded in polyethylene glycol (PEG) show good morphological preservation and increased detectability for immunocytochemical localization of antigenic sites, and semithin sections of Visio-Bond (VB) embedded tissue provide higher resolution of cellular structure, we applied these techniques and immunogold-silver stain (IGSS) for a more accurate localization of hepatic carbohydrate metabolic enzymes.

Sign in / Sign up

Export Citation Format

Share Document