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Optimal Preparation of Formalin Fixed Samples for Peptide Based Matrix Assisted Laser Desorption/Ionization Mass Spectrometry Imaging Workflows
08:16

Optimal Preparation of Formalin Fixed Samples for Peptide Based Matrix Assisted Laser Desorption/Ionization Mass Spectrometry Imaging Workflows

Published on: January 16, 2018

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Tissue fixed with formalin and processed without paraffin embedding is suitable for imaging of both peptides and

Monika Pietrowska1, Marta Gawin1,2, Joanna Polańska3

  • 1Center for Translational Research and Molecular Biology of Cancer, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology Gliwice Branch, Gliwice, Poland.

Proteomics
|March 23, 2016
PubMed

Insights

Formalin-fixed/frozen tissue offers a viable alternative for MALDI imaging. This method provides comparable peptide and lipid data to fresh-frozen and formalin-fixed/paraffin-embedded tissues, enhancing clinical compatibility.

Area of Science:

  • Biomedical Imaging
  • Analytical Chemistry
  • Histopathology

Background:

  • Sample preparation significantly impacts imaging mass spectrometry (IMS) outcomes.
  • Fresh-frozen tissues yield optimal IMS results but are not clinically practical.
  • Formalin-fixed paraffin-embedded (FFPE) tissues are standard in diagnostics but may alter molecular profiles.

Purpose of the Study:

  • To evaluate formalin-fixed tissue processed without paraffin embedding (formalin-fixed/frozen) for MALDI IMS of peptides and lipids.
  • To compare the utility of formalin-fixed/frozen samples against fresh-frozen and FFPE tissues.
  • To assess the compatibility of formalin-fixed/frozen samples with routine clinical workflows.

Main Methods:

  • MALDI imaging was performed on mouse kidney samples prepared as fresh-frozen, FFPE, and formalin-fixed/frozen.
  • Spectra from peptide and lipid imaging were compared across the different sample preparation methods.
  • Molecular images generated from formalin-fixed/frozen tissues were analyzed and compared to fresh-frozen and FFPE.

Main Results:

  • Peptide spectra from FFPE and formalin-fixed/frozen tissues showed high similarity.
  • Lipid spectra from fresh-frozen and formalin-fixed/frozen tissues exhibited strong resemblance.
  • Molecular images from formalin-fixed/frozen samples mirrored features of both fresh-frozen and FFPE for peptides, and fresh-frozen for lipids.

Conclusions:

  • Tissue preserved by formalin fixation and processed without paraffin embedding is a practical alternative for MALDI IMS.
  • This method bridges the gap between research-grade fresh-frozen tissues and clinically relevant FFPE samples.
  • Formalin-fixed/frozen tissue processing offers a promising approach for molecular imaging in clinical settings.

Keywords:
Clinical materialImaging mass spectrometryMouse kidneySample preparationTechnologyTissue fixation

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