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Related Experiment Videos

A sensitive double immunostaining protocol for Fos-immunoreactive neurons

Z C Peng1, S Chen, M Bentivoglio

  • 1Institute of Anatomy and Histology, University of Verona, Italy.

Brain Research Bulletin
|January 1, 1995
PubMed
Summary
This summary is machine-generated.

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This study introduces a novel double immunostaining technique for identifying activated neurons. The method uses sequential avidin-biotin-immunoperoxidase staining for sensitive detection of Fos protein and other cellular markers.

Area of Science:

  • Neuroscience
  • Cell Biology
  • Immunohistochemistry

Background:

  • Immunostaining of Fos protein is crucial for assessing neuronal activation.
  • Identifying the specific cell types expressing Fos requires double immunohistochemistry.
  • Existing methods may lack sensitivity or long-term storage capabilities.

Purpose of the Study:

  • To present a highly sensitive sequential two-color avidin-biotin-immunoperoxidase method for double immunostaining.
  • To enable simultaneous detection of Fos protein and other cellular antigens.
  • To facilitate long-term storage of immunostained tissue sections.

Main Methods:

  • A sequential two-color avidin-biotin-immunoperoxidase protocol was developed.
  • Metal-intensified diaminobenzidine (int-DAB) was used for dark brown/black Fos nuclear staining.

Related Experiment Videos

  • Alpha-naphthol/pyronin reaction yielded pink cytoplasmic staining for a second antigen.

    • Main Results:

    • The int-DAB and alpha-naphthol/pyronin combination provided high-contrast double immunostaining.
    • This method demonstrated superior contrast compared to int-DAB with conventional DAB staining.
    • The technique's sensitivity was confirmed in paradigms detecting Fos with p75 nerve growth factor receptor, parvalbumin, or calbindin D28k.

    Conclusions:

    • The described sequential immunostaining method is highly sensitive and effective for neuronal activation studies.
    • This protocol allows for the identification of specific neuronal populations expressing Fos.
    • The method supports long-term preservation of tissue sections for future analysis.