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Updated: Feb 14, 2026

Preparation of Whole Bone Marrow for Mass Cytometry Analysis of Neutrophil-lineage Cells
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Ruthenium counterstaining for imaging mass cytometry.

Raúl Catena1, Luis M Montuenga2, Bernd Bodenmiller1

  • 1Institute of Molecular Life Sciences, University of Zürich, Zürich, Switzerland.

The Journal of Pathology
|February 7, 2018
PubMed
Summary

Researchers developed a novel ruthenium tetroxide (RuO4) counterstain for imaging mass cytometry. This metal-based stain enhances visualization of tissue structures, improving the utility of this advanced imaging technique.

Keywords:
IMChigh-throughput histologyimaging mass cytometrymetal counterstainingnext-generation immunohistochemistryruthenium tetroxide

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Area of Science:

  • Biomedical Imaging
  • Pathology
  • Analytical Chemistry

Background:

  • Imaging mass cytometry (IMC) allows simultaneous detection of over 40 epitopes in tissues at subcellular resolution using metal tags.
  • Effective counterstaining is crucial in IMC to visualize overall tissue structure, as target antigens are often in rare cell populations.
  • Conventional counterstains like haematoxylin or fluorescent dyes are incompatible with IMC's metal-based detection.

Purpose of the Study:

  • To develop a novel metal-based counterstain compatible with imaging mass cytometry.
  • To enable visualization of general tissue architecture in IMC experiments.
  • To facilitate the routine application of IMC in research and clinical settings.

Main Methods:

  • Developed a ruthenium tetroxide (RuO4) based counterstaining method.
  • Utilized oxidation and covalent binding of tissue components to RuO4.
  • Applied the counterstain to tissue sections for IMC analysis.

Main Results:

  • The RuO4 counterstain effectively visualizes tissue structure in both cellular and stromal areas.
  • It provides contrast analogous to traditional histological stains like haematoxylin.
  • The method is compatible with IMC and other metal-based imaging techniques.

Conclusions:

  • A new RuO4 counterstain has been successfully developed for IMC.
  • This method provides essential structural context for IMC data.
  • The counterstain broadens the applicability of IMC in pathology and biomedical research.