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Related Experiment Videos

Signal amplification by combining two advanced immunohistochemical techniques.

D Russo1, A Ambrosino, A Vittoria

  • 1Department of Biological Structures, Functions and Technology, University of Naples Federico II, Italy.

European Journal of Histochemistry : EJH
|January 7, 2004
PubMed
Summary

A new combined immunohistochemical method (EVS/MICA) significantly enhances detection sensitivity and staining intensity for neuronal nitric oxide synthase, vasoactive intestinal polypeptide, and chromogranin A in animal tissues.

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Area of Science:

  • Immunohistochemistry
  • Neuroendocrinology
  • Veterinary Pathology

Background:

  • EnVision+ System (EVS) and Mirror Image Complementary Antibody (MICA) are sensitive immunohistochemical techniques.
  • Avidin-biotin complex (ABC) is a commonly used routine method for comparison.
  • Intestinal neuroendocrine cells and nerve structures are key targets in diagnostic pathology.

Purpose of the Study:

  • To compare the sensitivity of EVS and MICA immunohistochemical techniques.
  • To develop and evaluate a combined EVS/MICA method for enhanced antigen detection.
  • To assess the efficacy of these methods for identifying specific neuroendocrine markers in animal intestines.

Main Methods:

  • Comparison of EVS and MICA techniques against the standard ABC method.

Related Experiment Videos

  • Application of primary antibodies against neuronal nitric oxide synthase (nNOS), vasoactive intestinal polypeptide (VIP), and chromogranin A (Cr A).
  • Utilizing pig and sheep intestinal tissues fixed in Bouin and embedded in Paraplast.

    • Main Results:

    • The EVS method showed slightly superior sensitivity compared to MICA.
    • The combined EVS/MICA method demonstrated 4-8 times greater sensitivity and staining intensity than individual methods.
    • Clear identification of immunopositive structures was achieved at a primary antibody dilution of 1:256,000.

    Conclusions:

    • The combined EVS/MICA technique offers significantly improved sensitivity and staining intensity for immunohistochemistry.
    • This enhanced sensitivity is attributed to a higher concentration of revealing peroxidase molecules.
    • The combined method is highly recommended for detecting low-abundance tissue antigens, particularly nNOS, VIP, and Cr A.