scholarly journals APPLICATIONS OF FREEZE-SUBSTITUTION TO ELECTRON MICROSCOPE STUDIES OF INVERTEBRATE OOCYTES

1961 ◽  
Vol 9 (4) ◽  
pp. 785-798 ◽  
Author(s):  
Lionel I. Rebhun
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Freeze Substitution ◽  
Ice Crystal ◽  
Considerable Similarity ◽  
Golgi Bodies ◽  
Substitution Process ◽  
Present Technique ◽  
Lead Hydroxide ◽  
Inner Light

A modified freeze-substitution process is described which gives a low percentage (less than 5 per cent) of preparations of invertebrate eggs which appear to be ice crystal-free at the resolution of the electron microscope. The mitochondria show no membranes in these preparations but can be recognized by internal spaces with the size and the distribution of the cristae. The Golgi bodies resemble those seen with diffusion fixatives, but the limiting membranes are here double; that is, they appear to be a triple-layered sandwich with two outer dark approximately 25A layers and an inner light layer of the same thickness. The endoplasmic reticulum is clearly present and resembles that seen with diffusion fixatives. Here again, the limiting membranes are double with the same dimensions as those in the Golgi bodies. The membranes of the Golgi bodies and ER are seen after permanganate but not lead hydroxide staining. The hyaloplasm (or "cell sap") is crowded with 150 to 200A particles and these are also seen lining the ER membranes. In general, the structures as seen with the present technique show considerable similarity to those seen with diffusion fixatives.

Desiccation causes the proliferation of multicellular hairs, but not mucilage papillae, in Cryptothallus mirabilis (Hepatophyta): a correlated light and electron microscope study

1990 ◽  
Vol 68 (3) ◽  
pp. 697-706 ◽  
Author(s):  
Jeffrey G. Duckett ◽  
Karen S. Renzaglia ◽  
Keith Pell
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Acid Phosphatase ◽  
Endophytic Fungus ◽  
Electron Microscope Study ◽  
Secretory Phase ◽  
Golgi Bodies ◽  
Dense Cytoplasm ◽  
Thin Walled ◽  
Multicellular Hair

When Cryptothallus dries out over periods of 4–20 days, the dorsal surfaces of the thalli become covered with multicellular hairs. The distribution of mucilage papillae and the endophytic fungus are not affected by desiccation. The hairs are thin walled and highly vacuolated whereas the mucilage papillae, like their secretory counterparts in Marchantia and mosses, are thick walled with dense cytoplasm containing stacks of endoplasmic reticulum and numerous Golgi bodies. Cytochemistry shows that the secretion is rich in carbohydrates and is derived from Golgi vesicles. After an active secretory phase, senescence of the mucilage papillae is associated with acid phosphatase activity. Key words: Aneura, Cryptothallus, desiccation, liverwort, mucilage papilla, multicellular hair, ultrastructure.

The Effects of Calcium on the Ultrastructure of Cox's Orange Apples With Reference to Bitter Pit Disorder

1975 ◽  
Vol 23 (1) ◽  
pp. 55 ◽  
Author(s):  
HK Mahanty ◽  
BA Fineran
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Microscope Study ◽  
Electron Microscope Study ◽  
Thin Sections ◽  
Golgi Bodies ◽  
Cool Storage ◽  
Bitter Pit ◽  
Intermediate Results

An electron microscope study on thin sections of epidermal, hypodermal and cortical tissues has been made of calcium-sprayed and unsprayed apples in relation to bitter pit disorder. Golgi bodies, endoplasmic reticulum, mitochondria, chromoplasts, vacuoles and groundplasm were better preserved in sprayed apples during cool storage for up to 2 months than in unsprayed samples. In unsprayed apples these organelles were often drastically changed. The nucleus remained reasonably well preserved in all samples. Apples from trees sprayed only once gave somewhat intermediate results.

Light and electron microscope studies of cytoplasmic aggregates formed in barley cells in response to Erysiphe graminis

1976 ◽  
Vol 54 (14) ◽  
pp. 1647-1655 ◽  
Author(s):  
W. R. Bushnell ◽  
R. J. Zeyen
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Light Microscopy ◽  
Rough Endoplasmic Reticulum ◽  
Rapid Development ◽  
Epidermal Cells ◽  
Erysiphe Graminis ◽  
Synthetic Activity ◽  
Golgi Bodies

Cytoplasmic aggregates that formed in susceptible barley epidermal cells 11-12 h after inoculation with Erysiphe graminis were examined by light microscopy in living specimens and by electron microscopy in fixed specimens. Rapid development of the aggregate (5–10 min) suggested that cytoplasm migrated to the site of each aggregation. The aggregate contained features generally associated with areas of high metabolic and synthetic activity: abundant mitochondria, rough endoplasmic reticulum (associated with smooth cisternae), Golgi bodies, and polyribosomes. Leucoplasts and nuclei were sometimes near aggregates but not consistently. Microbodies and osmiophilic spherosomes were not present.

scholarly journals The secretion of the eggshell of Schistocerca gregaria: ultrastructure of the follicle cells during the termination of vitellogenesis and eggshell secretion

1980 ◽  
Vol 46 (1) ◽  
pp. 455-477
Author(s):  
S.J. Kimber
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Schistocerca Gregaria ◽  
Follicle Cells ◽  
Vitelline Membrane ◽  
Desert Locust ◽  
Golgi Bodies ◽  
Short Period ◽  
Distinct Morphology ◽  
Dense Product

The secretion of the eggshell by the follicle cells in the desert locust, Schistocerca gregaria, was studied using the electron microscope. The 3 layers of the eggshell, the vitelline membrane, the endochorion, and the exochorion, are produced in sequence over a short period of about 30–36 h. The follicle cells contain little rough endoplasmic reticulum (RER) and small inconspicuous Golgi bodies during vitellogenesis. As eggshell secretion approaches there is an increase in the amount of RER and Golgi cisternae contain electron-dense product. At each stage of the 3-phase secretion cycle the follicle cells contain Golgi bodies and secretion vesicles with distinct morphology. The follicle cells increase in breadth and decrease in height between the beginning and end of eggshell secretion. The endochorion ridges arise at the junction between follicle cells and appear to be moulded by the microvilli formed at this position. In the ovary prior to ovulation, the eggshell consists of a thin (0.5 micrometer) electron-dense vitelline and an outer fibrillar exochorion layer, 20–30 micrometer thick. Further changes take place in the vitelline membrane and the endochorion after oviposition, and a layer of curly fibres, the extrachorion, is secreted in the oviduct.

scholarly journals THE EFFECTS OF ENUCLEATION ON THE CYTOPLASMIC MEMBRANES OF AMOEBA PROTEUS

1968 ◽  
Vol 37 (2) ◽  
pp. 300-315 ◽  
Author(s):  
Charles J. Flickinger
Keyword(s):  
Endoplasmic Reticulum ◽  
Fine Structure ◽  
Electron Microscope ◽  
Golgi Apparatus ◽  
Amoeba Proteus ◽  
Granular Endoplasmic Reticulum ◽  
Whole Cells ◽  
Golgi Bodies ◽  
Cytoplasmic Membranes ◽  
In Cells

The dependence of cytoplasmic membranes upon the nucleus was studied by examining enucleated amebae with the electron microscope at intervals up to 1 wk after enucleation. Amebae were cut into two approximately equal parts, and the fine structure of the enucleated portions was compared with that of the nucleated parts and starved whole cells which had been maintained under the same conditions. Golgi bodies were diminished in size 1 day after enucleation and were not detected in cells enucleated for more than 2 days. The endoplasmic reticulum of enucleated cells appeared to increase in amount and underwent changes in its morphology. The sparsely scattered short tubules of granular endoplasmic reticulum present in unmanipulated amebae from stock cultures were replaced in 1–3-day enucleates by long narrow cisternae. In 3–7-day enucleates, similar cisternae of granular endoplasmic reticulum encircled areas of cytoplasm partially or completely. It was estimated that in most cases hundreds of these areas encircled by two rough membranes were formed per enucleated cell. The number of ribosomes studding the surface of the endoplasmic reticulum decreased progressively with time after enucleation. In contrast, the membranes of nucleated parts and starved whole cells did not undergo these changes. The possible identification of membrane-encircled areas as cytolysomes and their mode of formation are considered. Implications of the observations regarding nuclear regulation of the form of the Golgi apparatus and the endoplasmic reticulum are discussed.

High-pressure freezing of cells in suspension for low-voltage scanning electron microscopy (LVSEM)

1991 ◽  
Vol 49 ◽  
pp. 64-65
Author(s):  
Marek Malecki ◽  
James Pawley ◽  
Hans Ris
Keyword(s):  
Electron Microscopy ◽  
High Pressure ◽  
Low Voltage ◽  
Culture Media ◽  
Cell Structure ◽  
Freeze Substitution ◽  
Ice Crystal ◽  
High Pressure Freezing ◽  
Cell Culture Media ◽  
Voltage Scanning

The ultrastructure of cells suspended in physiological fluids or cell culture media can only be studied if the living processes are stopped while the cells remain in suspension. Attachment of living cells to carrier surfaces to facilitate further processing for electron microscopy produces a rapid reorganization of cell structure eradicating most traces of the structures present when the cells were in suspension. The structure of cells in suspension can be immobilized by either chemical fixation or, much faster, by rapid freezing (cryo-immobilization). The fixation speed is particularly important in studies of cell surface reorganization over time. High pressure freezing provides conditions where specimens up to 500μm thick can be frozen in milliseconds without ice crystal damage. This volume is sufficient for cells to remain in suspension until frozen. However, special procedures are needed to assure that the unattached cells are not lost during subsequent processing for LVSEM or HVEM using freeze-substitution or freeze drying. We recently developed such a procedure.

Copper Localization in Cirrhotic Rat Liver by Scanning Electron Microscopy

1969 ◽  
Vol 27 ◽  
pp. 38-39 ◽  
Author(s):  
J. C. Russ ◽  
E. McNatt
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Copper Acetate ◽  
Lead Citrate ◽  
Dense Bodies ◽  
Transmission Electron ◽  
Electron Mode ◽  
Scanning Electron

In order to study the retention of copper in cirrhotic liver, rats were made cirrhotic by carbon tetrachloride inhalation twice weekly for three months and fed 0.2% copper acetate ad libidum in drinking water for one month. The liver tissue was fixed in osmium, sectioned approximately 2000 Å thick, and stained with lead citrate. The section was examined in a scanning electron microscope (JEOLCO JSM-2) in the transmission electron mode.Figure 1 shows a typical area that includes a red blood cell in a sinusoid, a disse, and a portion of the cytoplasm of a hepatocyte which contains several mitochondria, peribiliary dense bodies, glycogen granules, and endoplasmic reticulum.

Ultrastructural comparison of prolactin adenomas of pituitary in men and women

1988 ◽  
Vol 46 ◽  
pp. 346-347
Author(s):  
R.C. Caughey ◽  
U.P. Kalyan-Raman
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Phosphate Buffer ◽  
Pituitary Adenomas ◽  
Osmium Tetroxide ◽  
Secretory Granules ◽  
Uranyl Acetate ◽  
En Bloc ◽  
Men And Women ◽  
Transmission Electron

Prolactin producing pituitary adenomas are ultrastructurally characterized by secretory granules varying in size (150-300nm), abundance of endoplasmic reticulum, and misplaced exocytosis. They are also subclassified as sparsely or densely granulated according to the amount of granules present. The hormone levels in men and women vary, being higher in men; so also the symptoms vary between both sexes. In order to understand this variation, we studied 21 prolactin producing pituitary adenomas by transmission electron microscope. This was out of a total of 80 pituitary adenomas. There were 6 men and 15 women in this group of 21 prolactinomas.All of the pituitary adenomas were fixed in 2.5% glutaraldehyde, rinsed in Millonig's phosphate buffer, and post fixed with 1% osmium tetroxide. They were then en bloc stained with 0.5% uranyl acetate, rinsed with Walpole's non-phosphate buffer, dehydrated with graded series of ethanols and embedded with Epon 812 epoxy resin.

High-pressure freezing and freeze substitution of plant tissue

1992 ◽  
Vol 50 (1) ◽  
pp. 748-749
Author(s):  
William P. Sharp ◽  
Robert W. Roberson
Keyword(s):  
High Pressure ◽  
Cell Structure ◽  
Leaf Tissue ◽  
Freeze Substitution ◽  
Crystal Formation ◽  
Ice Crystal ◽  
High Pressure Freezing ◽  
Cell Components ◽  
Cold Metal ◽  
Brome Grass

The aim of ultrastructural investigation is to analyze cell architecture and relate a functional role(s) to cell components. It is known that aqueous chemical fixation requires seconds to minutes to penetrate and stabilize cell structure which may result in structural artifacts. The use of ultralow temperatures to fix and prepare specimens, however, leads to a much improved preservation of the cell’s living state. A critical limitation of conventional cryofixation methods (i.e., propane-jet freezing, cold-metal slamming, plunge-freezing) is that only a 10 to 40 μm thick surface layer of cells can be frozen without distorting ice crystal formation. This problem can be allayed by freezing samples under about 2100 bar of hydrostatic pressure which suppresses the formation of ice nuclei and their rate of growth. Thus, 0.6 mm thick samples with a total volume of 1 mm3 can be frozen without ice crystal damage. The purpose of this study is to describe the cellular details and identify potential artifacts in root tissue of barley (Hordeum vulgari L.) and leaf tissue of brome grass (Bromus mollis L.) fixed and prepared by high-pressure freezing (HPF) and freeze substitution (FS) techniques.

Smooth endoplasmic reticulum in perfusion and immersion-fixed Leydig cells

1990 ◽  
Vol 48 (3) ◽  
pp. 82-83
Author(s):  
R.T.F. Bernard ◽  
R.H.M. Cross
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Steroid Hormones ◽  
Transmission Electron Microscope ◽  
Leydig Cells ◽  
Smooth Endoplasmic Reticulum ◽  
Random Mixture ◽  
Transmission Electron

Smooth endoplasmic reticulum (SER) is involved in the biosynthesis of steroid hormones, and changes in the organisation and abundance of this organelle are regularly used as indicators of changes in the level of steroidogenesis. SER is typically arranged as a meshwork of anastomosing tubules which, with the transmission electron microscope, appear as a random mixture of cross, oblique and longitudinal sections. Less commonly the SER appears as swollen vesicles and it is generally suggested that this is an artefact caused during immersion fixation or during immersion of poorly-perfused tissue.During a previous study of the Leydig cells of a seasonally reproducing bat, in which tissue was fixed by immersion, we noted that tubular SER and vesicular SER often occured in adjacent cells and sometimes in the same cell, and that the abundance of the two types of SER changed seasonally. We came to doubt the widelyheld dogma that vesicular SER was an artefact of immersion fixation and set out to test the hypothesis that the method of fixation does not modify the ultrastructure of the SER.

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