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Histological-Based Stainings Using Free-Floating Tissue Sections
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Conventional histological and cytological staining with simultaneous immunohistochemistry enabled by invisible

Larry E Morrison1, Mark R Lefever2, Heather N Lewis2

  • 1Roche Diagnostics Solutions (Ventana Medical Systems, Inc.), Tucson, AZ, 85755, USA. lemorrison73@gmail.com.

Laboratory Investigation; a Journal of Technical Methods and Pathology
|December 29, 2021
PubMed

Insights

This study introduces a novel method for simultaneous histological and immunohistochemistry staining on a single slide. This technique enhances diagnostic accuracy by allowing single-cell comparisons and conserving precious tissue specimens.

Area of Science:

  • Histopathology and Molecular Pathology
  • Biomedical Imaging and Instrumentation

Background:

  • Conventional histological stains (H&E) and immunohistochemistry (IHC) are crucial for diagnostics but require separate slides, limiting specimen use and single-cell analysis.
  • Current methods prevent direct comparison of H&E and IHC staining at the single-cell level due to different tissue sections.

Purpose of the Study:

  • To develop a method for performing conventional stains and IHC simultaneously on the same slide.
  • To enable direct comparison of staining results at the single-cell level and improve diagnostic efficiency.

Main Methods:

  • Utilized invisible IHC chromogens that do not interfere with conventional stains (H&E, mucicarmine, Papanicolaou).
  • Developed a dual-camera brightfield microscope for simultaneous visualization of both stain types.
  • Demonstrated the technique on formalin-fixed paraffin-embedded tissues and cytology specimens using UV and near-infrared absorbing chromogens.

Main Results:

  • Successfully achieved simultaneous H&E and IHC staining on single slides without visual interference.
  • Enabled direct, single-cell level comparison between conventional stains and IHC.
  • Extended the method to special stains and cytology, showcasing versatility.
  • Utilized multispectral imaging for quantitative analysis and flexible image production.

Conclusions:

  • This novel simultaneous staining technique overcomes limitations of separate slide methods, conserving tissue and enabling precise single-cell comparisons.
  • The dual-camera microscopy and invisible chromogen approach offers a powerful tool for enhanced histopathological diagnostics and research.
  • The method is adaptable to various conventional stains and specimen types, broadening its clinical and research applicability.

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