Ultrastructure and immunogold labeling of cytoplasmic fibrous bundles in tobacco leaf cells infected with tobacco vein mottling virus

1992 ◽  
Vol 50 (1) ◽  
pp. 676-677
Author(s):  
E. D. Ammar ◽  
D. W. Thombury ◽  
T. P. Pirone
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Immunogold Labeling ◽  
Infected Leaf ◽  
Transmission Electron ◽  
Aphid Vectors ◽  
Cylindrical Inclusions ◽  
Tobacco Vein Mottling Virus ◽  
Unknown Composition ◽  
Infected Tobacco

Tobacco vein mottling virus (TVMV) is a potyvirus, with flexuous filamentous particles ca. 765 nm long and 12-13 nm in diameter. TVMV is transmitted by sap inoculation and by aphid vectors. Cylindrical inclusions characteristic of infections with potyviruses have been found in the cytoplasm of TVMV-infected tobacco leaves and protoplasts, and the presence of fibrous inclusions of unknown composition in the cytoplasm and nuclei of TVMV-infected tobacco has been reported. Here, we report the results of transmission electron microscopy (TEM) and immunogold labeling of cytoplasmic fibrous bundles found in TVMV-infected leaf cells, providing evidence that these bundles are composed of TVMV virions and/or coat protein.

Immunogold labeling of HTLV-III/LAV in H9 cells studied by transmission electron microscopy

1986 ◽  
Vol 13 (3) ◽  
pp. 265-269 ◽  
Author(s):  
D. Pekovic ◽  
S. Garzon ◽  
H. Strykowski ◽  
D. Ajdukovic ◽  
M. Gornitsky ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Immunogold Labeling ◽  
Transmission Electron

scholarly journals Pitfalls of immunogold labeling: analysis by light microscopy, transmission electron microscopy, and photoelectron microscopy.

1987 ◽  
Vol 35 (8) ◽  
pp. 843-853 ◽  
Author(s):  
G B Birrell ◽  
K K Hedberg ◽  
O H Griffith
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Light Microscopy ◽  
Tannic Acid ◽  
Cultured Cells ◽  
Immunogold Labeling ◽  
Fish Gelatin ◽  
Nonspecific Binding ◽  
Gold Particles ◽  
Transmission Electron

The immunogold method is widely used to localize, identify, and distinguish cellular antigens. There are, however, some pitfalls that can lead to nonspecific binding, particularly in cytoskeletal studies with gold probes prepared from small gold particles. We present a list of suggestions for minimizing nonspecific binding, with particular attention to two problems identified in this study. First, we find that the method used to prepare the colloidal gold particles affects the degree of nonspecific binding. Second, the standard BSA-stabilized small gold probes evidently possess exposed regions that bind to the proteins of cytoskeletal preparations. This was investigated in whole-mount cytoskeletal preparations of cultured cells by use of light microscopy, transmission electron microscopy, and photoelectron microscopy of silver-enhanced specimens. Gold probes were made from approximately 5-nm particles generated by reduction of HAuCl4 with three different reducing agents: white phosphorus, sodium borohydride, and citrate-tannic acid. All three preparations stabilized in the conventional way showed significant levels of nonspecific binding, which was highest with citrate-tannic acid. This problem was largely solved with all three types of probes by including fish gelatin in the probe buffer, by substituting fish gelatin for the BSA stabilizer used to prepare the probes, or by pre-adsorption methods. Application of these techniques resulted in clear immunogold labeling patterns with minimal nonspecific background.

The influence of acetylsalicylic acid on oxylipin migration in Cryptococcus neoformans var.neoformansUOFS Y-1378

2008 ◽  
Vol 54 (2) ◽  
pp. 91-96 ◽  
Author(s):  
Olihile M. Sebolai ◽  
Carolina H. Pohl ◽  
Piet J. Botes ◽  
Pieter W.J. van Wyk ◽  
Johan L.F. Kock
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Cell Wall ◽  
Acetylsalicylic Acid ◽  
Cryptococcus Neoformans ◽  
Cell Walls ◽  
Bone Lesion ◽  
Acid Synthesis ◽  
Immunogold Labeling ◽  
Transmission Electron

In this paper we report the influence of acetylsalicylic acid on oxylipin migration in Cryptococcus neoformans var. neoformans UOFS Y-1378, previously isolated from human bone lesion. Transmission electron microscopy suggests that osmiophilic material originates in mitochondria and is deposited inside the yeast cell wall, from which it is excreted into the environment, along capsule protuberances, or through capsule detachments. Previous studies using immunogold labeling indicate that these osmiophilic layers contain 3-hydroxy oxylipins. In this study, the addition of acetylsalicylic acid (an inhibitor of mitochondrial function) in increasing amounts to the cells abrogated the migration of osmiophilic material, as well as capsule detachment from cell walls, and hence, oxylipin excretion. Consequently, we hypothesize that 3-hydroxy oxylipins are produced in mitochondria, probably via incomplete β-oxidation or fatty acid synthesis, from which they are deposited inside the cell wall and excreted through tubular protuberances attached to the surrounding capsules and (or) through detachment of these oxylipin-containing capsules.

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