A comparison of labelled antibody methods for the detection of virus antigens in cell monolayers

Oram, J.D.; Crooks, Alan J.

doi:10.1016/0022-1759(79)90023-1

Cited by 11 publications

(2 citation statements)

References 19 publications

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“…Irrespective of any variations in the embedding or labelling protocols to improve the labelling efficiency, the label density over the sections will always be low compared with the numbers of virus particles visible (van Lent & Verduin, 1986). The antigenic mass of viruses is low (Beesley, 1984) which in turn leads to the binding of few antibody molecules and thus low particle labelling (Oram & Crooks, 1979). Also, most of the virus particles seen in sections are below the surface and are unavailable for labelling (van Lent et al, 1990), and, in addition, there may be steric hindrance of the gold probes of any surface epitopes (Hyatt et al, 1988).…”

Section: Discussionmentioning

confidence: 99%

Statistical analysis of the distribution of gold particles over antigen sites after immunogold labelling

Glasbey

Roberts²

1997

Journal of Microscopy

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SummaryThe locations of immunogold-labelled particles on ultrathin sections of pellets of purified tobacco rattle virus were analysed statistically. The distributions were found to be inconsistent with a Poisson process. Instead, a physically based model, a modified Poisson process, is proposed and shown to fit the data well. The antigen sites to which antibodies are attached are assumed to follow a Poisson process and one gold particle sits on top of each site. However, if two sites are closer than a threshold then the physical size of the gold particles with attached antibodies pushes the particles apart, and if sites are nearer than a higher threshold there is a chance that surface tension will pull the gold particles together.

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Section: Discussionmentioning

confidence: 99%

Statistical analysis of the distribution of gold particles over antigen sites after immunogold labelling

Glasbey

Roberts²

1997

Journal of Microscopy

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“…Unfortunately good contrast and resolution are often achieved at the expense of antigenicity. This antigenic destruction is further compounded by the low antigenic mass of viruses and bacterial pili (Oram & Crooks, 1979). Each structure binds relatively few antibody molecules and immunolabelling with electron-dense markers is low.…”

Section: Introductionmentioning

confidence: 99%

The use of gold markers in immunocytochemical studies of microbiological organisms: a review

Beesley

1986

Journal of Microscopy

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SUMMARY Colloidal gold probes have revolutionized electron immunocytochemistry and are now used extensively for microbiological studies. The gold probes can be applied in one of several methods to gain valuable information from the sample. The pre‐embedding technique, the post‐embedding technique, the immunonegative stain technique, the immunoreplica technique and the immunofreeze‐fracture technique, are each described and their applications in microbiology, for diagnosis, for the detection of the site of antigen synthesis in host cells, for vaccine production and for genetic engineering are reported.

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The use of ultrathin cryosections for localisation of influenza virus antigens in infected vero cell cultures

Beesley

Campbell

1984

Histochemistry

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The localisation of influenza virus antigens in infected Vero cell monolayer cultures by post embedding immunoelectron microscopy requires both good resolution and the retention of antigenicity in the tissue sections. Ultrathin cryosections are superior to ultrathin resin sections for this purpose. The colloidal gold probe was used in conjunction with specific antibody preparations to localise three viral proteins. Antibody raised against haemagglutinin glycoproteins labelled the host cell membrane and the virus fringe without contamination of the host cell nucleus, whereas antibody raised against viral nuclear protein labelled throughout the host cell cytoplasm and nucleus. Matrix protein was localised within the nucleus and was associated with the host cell membrane of the infected cell. The appearance of all these proteins was maximal 24 h post infection.

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A comparison of labelled antibody methods for the detection of virus antigens in cell monolayers

Cited by 11 publications

References 19 publications

Statistical analysis of the distribution of gold particles over antigen sites after immunogold labelling

Statistical analysis of the distribution of gold particles over antigen sites after immunogold labelling

The use of gold markers in immunocytochemical studies of microbiological organisms: a review

The use of ultrathin cryosections for localisation of influenza virus antigens in infected vero cell cultures

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