Neuroscience and Behavioral Physiology

, Volume 48, Issue 2, pp 153–156 | Cite as

A Method of Enhanced Immunofluorescence by Amplification of the Immunoperoxidase Reaction

  • V. V. Gusel’nikovaEmail author
  • D. E. Korzhevskii

The aim of the present work was to develop a simple and effective means of converting the immunoperoxidase reaction into an immunofluorescence reaction. Use of fluorochrome-conjugated antibodies to horseradish peroxidase significantly increased the intensity of the immunofluorescence reaction as compared with the indirect Coons method, which was confirmed by quantitative evaluation of fluorescence intensity. The conversion method developed here does not require the protocol to be expanded by a large number of steps or any reagent substitutions, which facilitates the transition from the immunoperoxidase reaction to the immunofluorescent reaction.


immunoperoxidase reaction immunofluorescence reaction conversion amplification 


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  1. 1.
    V. V. Gusel’nikova, S. A. Bekoeva, V. F. Korzhevskaya, et al., “Histochemical and immunohistochemical identification of mast cells in the human myocardium,” Morfologiya, 147, No. 2, 80–86 (2015).Google Scholar
  2. 2.
    D. E. Korzhevskii, O. V. Kirik, M. N. Karpenko, et al., Theoretical Bases and Applied Use of Immunohistochemical Methods: Guidelines, SpetsLit, St. Petersburg (2012).Google Scholar
  3. 3.
    D. E. Korzhevskii (ed.), O. V. Kirik, E. S. Petrova, et al., Theoretical Bases and Applied Use of Immunohistochemical Methods: Guidelines, SpetsLit, St. Petersburg (2014), 2nd ed.Google Scholar
  4. 4.
    D. E. Korzhevskii, O. V. Kirik, E. G. Sukhorukova, et al., Molecular Morphology. Methods of Fluorescence and Confocal Laser Microscopy, SpetsLit, St. Petersburg (2014).Google Scholar
  5. 5.
    D. E. Korzhevskii, E. G. Sukhorukova, E. G. Gilerovich, et al., “Advantages and disadvantages of zinc-ethanol-formaldehyde as a fixer for immunocytochemical studies and confocal laser microscopy,” Morfologiya, 143, No. 2, 81–85 (2013).Google Scholar
  6. 6.
    E. G. Sukhorukova, “Nuclear protein NeuN in human substantia nigra neurons,” Morfologiya, 143, No. 2, 78–80 (2013).Google Scholar
  7. 7.
    E. G. Sukhorukova, O. V. Kirik, N. M. Zelenkova, and D. E. Korzhev skii, “Neuronal nuclear antigen NeuN – an indicator of the preservation of neural tissue and its suitability for immunohistochemical studies,” Med. Akad. Zh., No. 1, 63–67 (2015).Google Scholar
  8. 8.
    O. S. Alekseeva, V. V. Gusel’nikova, G. V. Beznin, D. E. Korzhevskii, “Prospects of the nuclear protein NeuN application as an index of functional state of the vertebrate nerve cells,” Zh. Evol. Biokhim. Fiziol., 51, No. 5, 313–323 (2015).PubMedGoogle Scholar
  9. 9.
    V. V. Gusel’nikova and D. E. Korzhevskii, “NeuN as a neuronal nuclear antigen and neuron differentiation marker,” Acta Naturae, 7, No. 2, 42–47 (2015).PubMedPubMedCentralGoogle Scholar
  10. 10.
    T. Ibaraki, M. Muramatsu, S. Takai, et al., “The relationship of tryptase-and chymase-positive mast cells to angiogenesis in stage I nonsmall cell lung cancer,” Eur. J. Cardiothorac. Surg., 28, No. 4, 617–621 (2005).CrossRefPubMedGoogle Scholar
  11. 11.
    G. Ranieri, M. Ammendola, R. Patruno, et al., “Tryptase-positive mast cells correlate with angiogenesis in early breast cancer patients,” Int. J. Oncol., 35, No. 1, 115–120 (2009).CrossRefPubMedGoogle Scholar
  12. 12.
    J. Vitte, “Human mast cell tryptase in biology and medicine,” Mol. Immunol., 63, No. 1, 18–24 (2015).CrossRefPubMedGoogle Scholar
  13. 13.
    G. Wang, C. L. Achim, R. L. Hamilton, et al., “Tyramide signal amplification method in multiple-label immunofluorescence confocal microscopy,” Methods, 18, No. 4, 459–464 (1999).CrossRefPubMedGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Laboratory for the Functional Morphology of the Central and Peripheral Nervous System, Department of General and Special MorphologyInstitute of Experimental MedicineSt. PetersburgRussia

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