Improved preservation of fine structure of deep-sea microorganisms by freeze-substitution after glutaraldehyde fixation

2011 ◽  
Vol 60 (4) ◽  
pp. 283-287 ◽  
Author(s):  
M. Yamaguchi ◽  
Y. Namiki ◽  
H. Okada ◽  
K. Uematsu ◽  
A. Tame ◽  
...  
Keyword(s):  
Fine Structure ◽  
Deep Sea ◽  
Freeze Substitution ◽  
Glutaraldehyde Fixation

Lipid in Erysiphe graminis hordei and its possible role during germination

1970 ◽  
Vol 16 (11) ◽  
pp. 1041-1044 ◽  
Author(s):  
W. E. McKeen
Keyword(s):  
Fine Structure ◽  
Electron Microscope ◽  
Osmium Tetroxide ◽  
Erysiphe Graminis ◽  
Glutaraldehyde Fixation ◽  
Sudan Black B ◽  
Erysiphe Graminis Hordei ◽  
Sudan Iv ◽  
Mother Cells ◽  
Hyphal Tip

Osmiophilic bodies appear in parts of the colonial growth of Erysiphe graminis DC. f. sp. hordei Em Marchal culture CR3 growing on the susceptible commercial Keystone variety of barley. They are readily observed by the light and electron microscope after osmium tetroxide staining and are abundant in conidiophores, conidia, and mycelium except in haustorial mother cells, in which they are usually absent. The metabolism of haustorial mother cells is distinct and the fine structure of adjoining cells is frequently different. Osmiophilic bodies are absent from the growing hyphal tip, but gradually increase in number and size further back in the terminal cell. Electron micrographs show that they are intracytoplasmic, intravacuolar, and up to 1 μ in diameter. When the colony is washed with acetone or alcohol rather than with aqueous buffer, after glutaraldehyde fixation, before osmium tetroxide fixation, the osmiophilic bodies are removed, indicating that they are lipids. Fat stains, Sudan black B, and Sudan IV stain these bodies. Perhaps the water needs of the germinating conidium are met in part by the oxidation of fats.

Physical Preservation Methods for Biological Material

1972 ◽  
Vol 30 ◽  
pp. 134-135
Author(s):  
Stanley Bullivant
Keyword(s):  
Freeze Drying ◽  
Freeze Substitution ◽  
Ice Crystals ◽  
Crystal Formation ◽  
Glycerol Solution ◽  
Ice Crystal ◽  
Glutaraldehyde Fixation ◽  
Preservation Methods ◽  
Glycerol Treatment ◽  
Pre Treatment

This review will be limited to physical preservation methods involving freezing, and will include discussion of freeze-drying, freeze-substitution, ultracryomicrotomy and freezeetching. Pre-treatment and freezing steps can be considered as common to all techniques.The most common pre-treatment is soaking in a cold 20% glycerol solution to provide protection against ice crystal formation. Glutaraldehyde fixation is often used before freezing. With special freezing techniques no pretreatment may be necessary.Freezing should be so rapid that it does not produce ice crystals larger than 2nm. in diameter. Following glycerol treatment, immersion in Freon 12 at its melting point of -155 C is adequate. Freezing in boiling liquid nitrogen is not usually satisfactory.

scholarly journals Possible continuity of subplasmalemmal cytoplasmic network with basement membrane cord network: ultrastructural study

1995 ◽  
Vol 108 (5) ◽  
pp. 1971-1976
Author(s):  
S. Inoue
Keyword(s):  
Basement Membrane ◽  
Type Iv Collagen ◽  
Freeze Substitution ◽  
High Resolution Electron Microscopy ◽  
Sulfate Proteoglycan ◽  
Glutaraldehyde Fixation ◽  
Lamina Densa ◽  
Type Iv ◽  
Resolution Electron ◽  
Visceral Epithelium

The ultrastructure of the subplasmalemmal cytoplasm of the cell and the associated basement membrane as well as the area of the cell-basement membrane border were observed with high resolution electron microscopy after preparation of the tissues with cryofixation or glutaraldehyde fixation followed by freeze substitution. The subplasmalemmal cytoplasm of the smooth muscle cells of rat epididymal tubules and the podocyte processes of the mouse glomerular visceral epithelium were found to be composed of a fine network of irregular anastomosing strands. This network closely resembled the previously characterized cord network of the basement membrane. The cords are known to be composed of a 1.5 to 3 nm thick core filament made up of type IV collagen which is surrounded by an irregular ‘sheath’ of other components. The strands in the subplasmalemmal network showed ultrastructural features similar to those of the cord network. Ribbon-like, 4.5 nm wide heparan sulfate proteoglycan ‘double tracks’ were previously reported to be associated with the cord network. Structures similar in size and appearance to the double tracks were also found in the subplasmalemmal network. At the cell-basement membrane border, the lamina densa of the basement membrane was in contact with the cell without the intervening space of a lamina lucida which was recently found to be an artefact caused by conventional tissue processing. Furthermore, the subplasmalemmal network appeared to be continuous through the plasma membrane, with the cord network of the basement membrane.(ABSTRACT TRUNCATED AT 250 WORDS)

Fine structure of peripheral terminations in the porous sensillar cone of larvae of Ctenicera destructor (Brown) (Coleoptera, Elateridae), and probable fixation artifacts

1971 ◽  
Vol 49 (6) ◽  
pp. 789-799 ◽  
Author(s):  
R. Y. Zacharuk
Keyword(s):  
Fine Structure ◽  
Glutaraldehyde Fixation ◽  
Dendritic Branches ◽  
Peripheral Sensory ◽  
Ammoniacal Silver ◽  
Ctenicera Destructor

Nodes occur naturally on the dendritic branches in the receptor-lymph cavity of the antennal sensory cone in larvae of Ctenicera destructor. They increase in number, are often greatly expanded, and may separate from the dendrites as fixation artifacts. Extracellular dictyosomes originate from terminal nodes of the dendritic branches. These dictyosomes secrete vesicles, believed to contain lipoidal substances, directly into the fluid in the receptor cavity. These vesicles either coalesce into large lipoidal globules which produce the peripheral sensory tubules that traverse pores in the covering cuticle, or themselves attach directly to the tubules. The sensory tubules are lipoidal in nature. It is suggested that they serve to conserve moisture in the sensillum and to channel stimulating molecules through the cuticle to the lymph in the receptor cavity. The dendrites are not attached directly to the sensory tubules, but are believed to gain contact with the stimulating molecules through the receptor lymph. The lipoidal substance for sensory tubule secretion is probably transported into the sensory cone by the neurotubules within the dendrites. The results from a technique using glutaraldehyde fixation, ammoniacal silver treatment, and lead staining, without osmium fixation, are described and discussed.

scholarly journals SARCOLEMMAL INVAGINATIONS CONSTITUTING THE T SYSTEM IN FISH MUSCLE FIBERS

1964 ◽  
Vol 22 (3) ◽  
pp. 675-696 ◽  
Author(s):  
Clara Franzini-armstrong ◽  
Keith R. Porter
Keyword(s):  
Fine Structure ◽  
Osmium Tetroxide ◽  
Striated Muscle ◽  
Muscle Fibers ◽  
Fish Muscle ◽  
Structural Features ◽  
The Body ◽  
Glutaraldehyde Fixation ◽  
Striated Muscle Fibers ◽  
T System

Striated muscle fibers from the body and tail myotomes of a fish, the black Mollie, have been examined with particular attention to the sarcoplasmic reticulum (SR) and transverse tubular (or T) system. The material was fixed in osmium tetroxide and in glutaraldehyde, and the images provided by the two kinds of fixatives were compared. Glutaraldehyde fixes a fine structure that is broadly comparable with that preserved by osmium tetroxide alone but differs in some significant details. Especially significant improvements were obtained in the preservation of the T system, that is, the system of small tubules that pervades the fiber at every Z line or A-I junction level. As a result of this improved glutaraldehyde fixation, the T system is now clearly defined as an entity of fine structure distinct from the SR but uniquely associated with the SR and myofibrils. Glutaraldehyde fixation also reveals that the T system is a sarcolemmal derivative that retains its continuity with the sarcolemma and limits a space that is in direct communication with the extracellular environment. These structural features favor the conclusion that the T system plays a prominent role in the fast intracellular conduction of the excitatory impulse. The preservation of other elements of muscle fine structure, including the myofibrils, seems for reasons discussed, to be substantially improved by glutaraldehyde.

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