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Updated: Jan 26, 2026

Expanding the Comprehension of the Tumor Microenvironment using Mass Spectrometry Imaging of Formalin-Fixed and Paraffin-Embedded Tissue Samples
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Optimal Formalin-Fixed Paraffin-Embedded Sample Preservation for Efficient Staining in Multiplex Imaging.

Yuna Delarue1, Nadège Marec2, Marion Le Rochais1

  • 1Centre Hospitalier Universitaire de Brest, Brest, France; LBAI, UMR1227, Univ Brest, Inserm, Brest, France.

Laboratory Investigation; a Journal of Technical Methods and Pathology
|January 24, 2026
PubMed
Summary
This summary is machine-generated.

Room temperature storage degrades immunohistochemical signals over time, impacting diagnostic accuracy. Storing tissue sections at -20°C or below preserves signal integrity for reliable analysis.

Keywords:
Hyperionhistologic slideimaging mass cytometrypreservationstorage

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Area of Science:

  • Biopathology
  • Immunohistochemistry
  • Biomarker Analysis

Background:

  • Immunohistochemistry accuracy relies on preserving antigenic integrity in tissue samples.
  • Formalin-fixed paraffin-embedded tissues are common for storage, but signal degradation over time is a concern.
  • The impact of storage conditions on marker detection remains poorly understood.

Purpose of the Study:

  • To investigate the extent of signal degradation in stored tissue samples.
  • To evaluate the influence of different storage conditions on marker detection.
  • To assess the effectiveness of normalization techniques for preserving data quality.

Main Methods:

  • Analysis of tissue blocks and sections stored for up to eight years under various conditions (room temperature, 4°C, -20°C, -80°C).
  • Staining with 25 antibodies targeting immune, stromal, and structural markers using imaging mass cytometry (IMC).
  • Quantification and analysis of signal intensities, with and without normalization via the imcRtools pipeline.

Main Results:

  • Gradual signal loss observed in room temperature-stored blocks (>6 years), affecting specific markers.
  • Significant signal degradation occurred within weeks at room temperature for sensitive markers like CD20, CD45, and CD45RA.
  • -20°C or -80°C storage maintained staining quality; 4°C offered partial preservation.
  • Normalization partially corrected variations but could not recover severely degraded signals.

Conclusions:

  • Room temperature storage significantly reduces immunoreactivity detectable by IMC, potentially affecting scientific interpretation and diagnostics.
  • Storing tissue sections at -20°C is an effective and accessible method for preserving immunohistochemical quality.
  • For retrospective studies, prioritize tissue blocks stored for less than six years; standardized preservation protocols are crucial for reproducibility.