Immunocytochemistry (Up to date 2015)

Author: Susan Van Noorden

Preservation of Cytological Samples for Immunocytochemistry

Foreword

Immunocytochemistry is now essential for full and accurate pathological diagnosis of tissue samples whether of formalin-fixed blocks or cell preparations from brushings, scrapings, smears or fine needle aspirates. The success of the method depends to a large extent on the morphological preservation of the cells as well as of the antigens to be identified. Poorly made or unsuitably preserved smears or other preparations waste the sample, reagents and time, only to give unreliable results. Therefore the importance of the initial treatment of the material is paramount.

Contents

  • Preparation and fixation of samples

  • Principles of Immunocytochemistry

    • Antigen retrieval

    • Blocking endogenous peroxidase

    • Blocking non-specific background

  • Antibody dilution

    • Diluent                                                                                                                           

    • Optimisation

  • Controls

  • Apparatus

  • Special considerations for immmunocytopathology

  • Appendix

    • Esposti’s fluid

    • Buffers

      • Phosphate-buffered saline

      • Tris-buffered saline

    • Antibody diluent

    • Antibody storage

    • Blocking endogenous peroxidase

    • Heat-induced epitope retrieval

      • Citrate buffer

      • TRIS/EDTA buffer

      • microwaving method

    • Diaminobenzidine solution for peroxidase development

    • Immunostaining method (manual)

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