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Combined Immunohistochemistry after Mass Spectrometry Imaging for Superior Spatial Information

Katharina Kriegsmann1, Rémi Longuespée2, Michael Hundemer1

  • 1Department of Hematology, Oncology and Rheumatology, University of Heidelberg, 69117, Heidelberg, Germany.

Proteomics. Clinical Applications
|July 24, 2018
PubMed

Insights

Immunohistochemistry (IHC) is feasible on tissue slides after Mass Spectrometry Imaging (MSI), though staining intensity may decrease. This allows for better cell identification in complex samples like bone marrow and lung cancer tissues.

Area of Science:

  • Pathology
  • Biomedical Imaging
  • Cancer Research

Background:

  • Hematoxylin and Eosin (H&E) staining aids region identification in Mass Spectrometry Imaging (MSI) but can limit specific cell identification.
  • Immunohistochemistry (IHC) enhances cell visualization but its application on MSI-analyzed tissues is unexplored.

Purpose of the Study:

  • To investigate the feasibility and efficacy of single and combined IHC staining on tissue sections previously analyzed by MSI.
  • To evaluate the impact of MSI on IHC staining quality and intensity.

Main Methods:

  • MSI was performed on bone marrow biopsies (multiple myeloma) and lung cancer tissues (adenocarcinoma, squamous cell carcinoma).
  • Specific antibodies (e.g., CD38, CD138, MUM1, kappa, lambda, CK5/6/TTF1, Napsin-A/p40) were used for IHC staining post-MSI.
  • Staining intensities of p40 were digitally quantified and compared between MSI-analyzed sections and serial sections using a tissue microarray (n=44).

Main Results:

  • IHC staining, including single and double combinations, was found to be feasible on tissue sections analyzed by MSI.
  • A decrease in staining intensity was observed after MSI compared to serial sections.
  • Digital analysis confirmed that despite reduced intensity, the staining quality and quantity were sufficient for identifying cells of interest.

Conclusions:

  • Single and double IHC staining can be successfully applied to tissue sections following MSI analysis.
  • While MSI affects staining intensity, it does not preclude the identification of target cells, offering a valuable combined approach for complex tissue analysis.
Abstract

Keywords:
MSMS imagingdigital image analysisimmunohistochemistrymolecular pathology

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