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Immunogold cytochemistry in neuroscience

Nature Neuroscience volume 16, pages 798–804 (2013)Cite this article

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Abstract

The complexity of the central nervous system calls for immunocytochemical procedures that allow target proteins to be localized with high precision and with opportunities for quantitation. Immunogold procedures stand out as particularly powerful in this regard. Although these procedures have found wide application in the neuroscience community, they present limitations and pitfalls that must be taken into account. At the same time, these procedures offer potentials that remain to be fully realized.

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Figure 1: Principles underlying a two-step, postembedding immunogold procedure.
Figure 2: Experimental determination of anatomical resolution.
Figure 3: Gold particles signaling a membrane protein epitope are normally distributed around the plasma membrane.
Figure 4: Targeted disruption of an anchoring molecule confirms selectivity of antibody under the conditions of immunocytochemistry.
Figure 5: Positive control, showing that the antibody recognizes the target antigen under the conditions of immunocytochemistry.
Figure 6: Positive control, validating immunocytochemical procedure for small molecular antigens.
Figure 7: Sandwiches like those used for validation can be used to establish calibration curves.
Figure 8: Combining immunogold labeling for glutamate receptors and glutamate.

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Acknowledgements

We thank G. Lothe and C. Knudsen for technical assistance, P. Laake for comments, and D. Frydenlund for providing Figure 5. We gratefully acknowledge support from the Norwegian Research Council, the European Union framework programmes and the Letten Fund.

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  1. Laboratory of Molecular Neuroscience, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway

    Mahmood Amiry-Moghaddam & Ole Petter Ottersen

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Correspondence to Ole Petter Ottersen.

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Amiry-Moghaddam, M., Ottersen, O. Immunogold cytochemistry in neuroscience. Nat Neurosci 16, 798–804 (2013). https://doi.org/10.1038/nn.3418

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