Double Immunohistochemistry and Digital Image Analysis

Methods Mol Biol. 2019:1913:3-11. doi: 10.1007/978-1-4939-8979-9_1.

Abstract

Immunohistochemistry (IHC) is a commonly used technique for protein detection in tissue sections. The method requires high-affinity antibodies that are specific for the target proteins of interest. More advanced IHC techniques have been developed to meet the need for simultaneous detection of more than one target protein in the same tissue section. This chapter provides general guidelines for double IHC staining of formalin-fixed, paraffin-embedded tissue sections. Chromogenic substrates are chosen based on their excellent contrast and compatibility with the subsequent digital image analysis.

Keywords: Antibody; Chromogenic substrate; Digital image analysis; Double immunohistochemistry; FFPE.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD34 / analysis
  • Biomarkers, Tumor / analysis
  • Fluorescent Antibody Technique / instrumentation
  • Fluorescent Antibody Technique / methods*
  • Humans
  • Image Processing, Computer-Assisted / instrumentation
  • Image Processing, Computer-Assisted / methods*
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods
  • Neoplasms / pathology*
  • Paraffin Embedding / instrumentation
  • Paraffin Embedding / methods
  • Receptor, Platelet-Derived Growth Factor beta / analysis
  • Software
  • Tissue Fixation / instrumentation
  • Tissue Fixation / methods

Substances

  • Antigens, CD34
  • Biomarkers, Tumor
  • PDGFRB protein, human
  • Receptor, Platelet-Derived Growth Factor beta