Immunoelectron Microscopy for Virus Identification

Milne, Robert G.

doi:10.1007/978-3-642-75818-8_7

Cited by 16 publications

(10 citation statements)

References 65 publications

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“…IEM was performed as described with a few modifications [17–18]. Briefly, cells were fixed with 4% paraformaldehyde and 1% glutaral in 0.1 M phosphate buffer at 4°C for 4 hrs.…”

Section: Methodsmentioning

confidence: 99%

Secretion of annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients

Yin

Yan

Yao

et al. 2012

J Cellular Molecular Medi

106

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Early detection of resistance to platinum-based therapy is critical for improving the treatment of ovarian cancers. We have previously found that increased expression of annexin A3 is a mechanism for platinum resistance in ovarian cancer cells. Here we demonstrate that annexin A3 can be detected in the culture medium of ovarian cancer cells, particularly these cells that express high levels of annexin A3. Levels of annexin A3 were then determined in sera from ovarian cancer patients using an enzyme-linked immunosorbent assay. Compared with those from normal donors, sera from ovarian cancer patients contain significantly higher levels of annexin A3. Furthermore, serum levels of annexin A3 were significantly higher in platinum-resistant patients than in platinum-sensitive patients. To gain insight into the mechanism of secretion, the ovarian cancer cell lines were examined using both transmission electron microscopy and immunoelectron microscopy. Compared with parent cells, there are significantly more vesicles in the cytoplasm of ovarian cancer cells that express high levels of annexin A3, and at least some vesicles are annexin A3-positive. Moreover, some vesicles appear to be fused with the cell membrane, suggesting that annexin A3 secretion may be associated with exocytosis and the release of exosomes. This is supported by our observation that ovarian cancer cells expressing higher levels of annexin A3 released increased numbers of exosomes. Furthermore, annexin A3 can be detected in exosomes released from cisplatin-resistant cells (SKOV3/Cis) by immunoblotting and immunoelectron microscopy.

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“…IEM was performed as described with a few modifications [17–18]. Briefly, cells were fixed with 4% paraformaldehyde and 1% glutaral in 0.1 M phosphate buffer at 4°C for 4 hrs.…”

Section: Methodsmentioning

confidence: 99%

Secretion of annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients

Yin

Yan

Yao

et al. 2012

J Cellular Molecular Medi

106

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“…Virus identification was carried out by using the immunoelectron-microscopical method of decoration with antisera (Milne and Luisoni, 1977 For serological investigations standard DAS-ELISA as described by Clark and Adams (1977) and the plate trapped antigen (PTA)-ELISA according to Richter et al (1994) were carried out. Sample preparation and SDS-PAGF for "Western blotting were used as described by Laemmli (1970).…”

Section: Methodsmentioning

confidence: 99%

Evidence of a plant virus in Iceland

Kegler

Ehrig

Rabenstein

1998

Archives Of Phytopathology And Plant Protection

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In the south-western part of Iceland virus-like symptoms were observed on various plant species as Betula pubescens, Epilobium angustifolium, Geranium syhatkum, Lupinusnootkatensis, Paeonia peregrina, Sorbus ancuparia, Stachys betonicifolia. A mechanically transmissible virus could be isolated from E. angustifolium. By investigating the host range and by immunoelectron-microscopical and serological methods the isolated virus was identified as turnip mosaic potyvirus. Thus, a plant pathogenic virus was detected in Iceland for the first time.

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“…To improve the attachment of virus particles, grids can be coated by an antiserum prior to incubation with the leaf extract. This ‘trapping’ of particles results in a selective immune‐enrichment of virus particles on the grid (Milne, ). Another advantage of the layer of antibodies and other serum constituents is the inhibition of non‐specific binding of structures from the sample preparation.…”

Section: General Instructionsmentioning

confidence: 99%

“…Following incubation and washing, the grid can either be negatively stained and observed or a secondary antibody (which may be conjugated to gold particles) can be used prior to negative staining. In this case, the sample is incubated with a drop of diluted anti‐virus antiserum, so that antibodies can attach to the virus particles (Milne, ). Decoration with homologous antibodies (IEM) followed by staining will usually allow virus identification, provided that the analytical specificity of the antibodies is known.…”

Section: General Instructionsmentioning

confidence: 99%

PM 7/126 (1) Electron microscopy in diagnosis of plant viruses

2015

EPPO Bulletin

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Immunoelectron Microscopy for Virus Identification

Cited by 16 publications

References 65 publications

Secretion of annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients

Secretion of annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients

Evidence of a plant virus in Iceland

PM 7/126 (1) Electron microscopy in diagnosis of plant viruses

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