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Updated: Sep 19, 2025

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Label-Free Immunohistochemistry Imaging by Oblique-Incidence Reflectivity Difference

Meng Li1, Yuanyuan Xie1, Dandan Ji1

  • 1State Key Laboratory of Resource Insects; Key Laboratory of Luminescence Analysis and Molecular Sensing (Southwest University), Ministry of Education; School of Materials and Energy, Southwest University, Chongqing 400715, P. R. China.

Analytical Chemistry
|June 5, 2025
PubMed

Insights

This study introduces a novel label-free immunohistochemistry (IHC) method using oblique-incidence reflectivity difference (OIRD) imaging. This technique maps biomarker distribution without labeled antibodies, advancing pathological analysis and disease diagnosis.

Area of Science:

  • Biomedical Imaging
  • Pathology
  • Biochemistry

Background:

  • Immunohistochemistry (IHC) is a standard technique in pathology for visualizing biomarkers in tissues.
  • Conventional IHC relies on labeled antibodies, which can be costly and time-consuming.
  • There is a need for more efficient and accessible methods for biomarker detection.

Purpose of the Study:

  • To develop and validate a label-free IHC imaging strategy.
  • To demonstrate the capability of oblique-incidence reflectivity difference (OIRD) imaging for biomarker mapping.
  • To assess the potential of this method for pathological analysis and drug screening.

Main Methods:

  • Utilized oblique-incidence reflectivity difference (OIRD) imaging technique.
  • Applied OIRD to tissue slices for label-free biomarker visualization.
  • Monitored dynamic antibody binding kinetics.
  • Validated results against traditional fluorescent IHC.

Main Results:

  • Successfully mapped the spatial distribution of biomarkers without labeled antibodies.
  • Demonstrated OIRD imaging's efficacy in label-free IHC.
  • Observed and monitored antibody-tissue binding kinetics.
  • Confirmed the strategy's effectiveness through comparison with fluorescent IHC.

Conclusions:

  • Introduced a pioneering label-free IHC imaging strategy using OIRD.
  • This method obviates the need for labeled antibodies, simplifying the process.
  • The OIRD technique offers a promising tool for pathological analysis, disease diagnosis, and drug screening.

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