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Journal of Molecular Histology

, Volume 39, Issue 4, pp 459–462 | Cite as

A simple method for the production of bacterial controls for immunohistochemistry and fluorescent in situ hybridization

  • Camilla Recordati
  • Enrico Radaelli
  • Kenneth W. Simpson
  • Eugenio Scanziani
Brief Communication

Abstract

Immunohistochemical and fluorescent in situ hybridization (FISH) assays are useful diagnostic methods for the identification of bacteria on formalin fixed paraffin embedded histological sections. To validate an anti-bacterial antibody or an oligonucleotide probe and to ensure fidelity during subsequent analyses, suitable positive and negative controls are necessary. Suspensions of fixed bacteria are often used, but ideally, these controls should be fixed, embedded and processed in the same way of tissue samples under analysis. Herein, we describe a simple method for the production of bacterial histological control samples: the sandwich. The sandwich is composed of two external layers of equine lung parenchyma and a central layer of the target bacterium. We prepared sandwiches containing Escherichia coli, Campylobacter jejuni, and Arcanobacterium pyogenes and tested them with appropriate antibodies and Eub338 FISH probe. The sandwich is an effective and simple method to prepare bacterial histological controls fixed and processed in the same way as the diagnostic tissues.

Keywords

Immunohistochemistry Fluorescent in situ hybridization Bacteria Controls 

Notes

Acknowledgements

We thank A. Invernizzi for the bacterial cultures, G. Lombardi for the rabbit polyclonal anti-A. pyogenes antibody, and M. Barzani and M. Brevi for technical support.

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Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Camilla Recordati
    • 1
  • Enrico Radaelli
    • 1
  • Kenneth W. Simpson
    • 2
  • Eugenio Scanziani
    • 1
  1. 1.Dipartimento di Patologia Animale, Igiene e Sanità Pubblica Veterinaria, Sezione di Anatomia Patologica Veterinaria e Patologia Aviare, Facoltà di Medicina VeterinariaUniversità degli Studi di MilanoMilanoItaly
  2. 2.Department of Clinical Sciences, College of Veterinary MedicineCornell UniversityIthacaUSA

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