Multiple biotin-avidin amplification for multiple immunostaining

Gian Luca Ferri, Rosa Maria Gaudio, Filippa Ida Castello, Cataldo Tirolo, Francesca Chiolerio

Research output: Contribution to journalArticlepeer-review


A multiple fluorescence immunostaining method is described, based on the use of multiple fluorochrome-avidin/streptavidin conjugates. Key factors are (a) stabilization of each (preceding) biotin-avidin chain and associated avidin blocking step with formaldehyde and (b) application of two or three (possibly more) fluorochrome-labeled avidins/streptavidins in decreasing order of visual sensitivity. Under the conditions described, no cross-talk was detected, and both signal and background levels were comparable to corresponding single immunostaining. Enzyme-linked immunosorbent assay experiments confirmed effective inhibition of competition-displacement of previously bound avidin after the blocking-stabilization procedure used.

Original languageEnglish
Pages (from-to)73-80
Number of pages8
JournalApplied Immunohistochemistry
Issue number1
Publication statusPublished - Mar 1999


  • Avidin
  • Biotin
  • Double immunostaining
  • Fluorochrome
  • Immunocytochemistry
  • Immunofluorescence
  • Immunostaining
  • Multiple immunostaining

ASJC Scopus subject areas

  • Anatomy
  • Medical Laboratory Technology


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