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

Beesley, Julian E.

doi:10.1111/j.1365-2818.1986.tb02776.x

Cited by 13 publications

(2 citation statements)

References 10 publications

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“…This is perhaps one of the easiest of all the immunocytochemical techniques (Beesley, 1986). Crude membrane preparations (Fig.…”

Section: Negative Staining and Immunogold Labellingmentioning

confidence: 99%

Immunocytochemical strategies for electron microscopy: choice or compromise

Skepper

2000

Journal of Microscopy

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SummaryThis review attempts to describe the major immunocytochemical strategies that are in current use for transmission electron microscopy. Emphasis is placed on suggesting the most appropriate application of each technique that is described. The review is not intended to identify the microscopist's`cure all' technique for immunocytochemistry but rather to describe what techniques are available and what are their relative merits. Expensive equipment and technically complicated methods are not always necessary to collect robust data. I hope to present a review that will enable its readers to select the most profitable technique (or compromise) available, to reflect both their resources and their ability to address their biological questions with vigour.

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“…This is perhaps one of the easiest of all the immunocytochemical techniques (Beesley, 1986). Crude membrane preparations (Fig.…”

Section: Negative Staining and Immunogold Labellingmentioning

confidence: 99%

Immunocytochemical strategies for electron microscopy: choice or compromise

Skepper

2000

Journal of Microscopy

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“…Colloidal gold particles, conjugated to antibodies or other proteins, have proved to be good markers for the light and electron microscopic detection and quantification of antigens in many biological samples (Roth, 1983; Bendayan, 1985, 2000; Horisberger, 1985; Beesley, 1986). In this paper we introduce the use of colloidal gold probes for a solid phase, transmission electron microscope‐based immunodetection method for prior detection of antigen in cells using ultra low volumes of sample and reagents.…”

Section: Introductionmentioning

confidence: 99%

Pre‐screening for antigen detectability in cells: a TEM‐based solid phase digital immunogold detection method utilizing ultra low volumes of reagents

Kaur

Dikshit

Raje

2002

Journal of Microscopy

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SummaryRapid and sensitive pre-screening for the presence of antigens in cell samples and confirmation of reactivity of antibodies, before proceeding with electron microscopy, is highly desirable. Most of the methods developed for this purpose are generally not very efficient and suitable for dealing with very small volumes of sample and reagents. In this work we present a simple, sensitive and rapid solid phase transmission electron microscope (TEM) based method for the detection of picogram (pg) levels of soluble antigens using as little as 10 µ L of reagents. Protein was adsorbed onto grids coated with polystyrene films to form the solid phase. The presence of antigen was detected using immunogold labelling. Gold particles adhering to the film were visualized and counted in a TEM providing a digital signal. This method was 100-fold more sensitive than dot blot in detection of rabbit IgG. We have demonstrated the utility of this technique by screening for Vitreoscilla haemoglobin (VHb) antigen in cell lysates and confirming the results directly with immunogold labelling transmission electron microscopy of cell sections.

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The use of monoclonal antibodies with colloidal gold-labeled probes in postembedding immunoelectron microscopy

Suárez

Brown

1997

Mol Biotechnol

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This article focuses on procedures for the use of monoclonal antibodies (MAb) in immunoelectron microscopy (IEM) for the subcellular localization of antigens or to relate function with structure in prokaryotic and eukaryotic cells using postembedding immunoelectron microscopy techniques. The use of MAbs greatly increases the specificity and quality of information when used in combination with gold-labeled probes. Because of its specificity, the reactivity of MAb may be very sensitive to antigenic changes resulting from the process of sample preparation when performing IEM studies. Specific protocols for each particular combination of epitope/MAb must be usually specifically devised, since it is impossible to predict an experimental system that will successfully preserve structures and antigenic determinants for every combination. In this article, we discuss critical technical aspects that usually result in improved resolution when the procedure is used to identify structure in diverse prokaryotic and eukaryotic cells.

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

Cited by 13 publications

References 10 publications

Immunocytochemical strategies for electron microscopy: choice or compromise

Immunocytochemical strategies for electron microscopy: choice or compromise

Pre‐screening for antigen detectability in cells: a TEM‐based solid phase digital immunogold detection method utilizing ultra low volumes of reagents

The use of monoclonal antibodies with colloidal gold-labeled probes in postembedding immunoelectron microscopy

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