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Impact of Tissue Sample Preparation Methods on Myelin-Sensitive Quantitative MR Imaging

Amaya Murguia1, Scott D Swanson2,3, Ulrich Scheven4

  • 1Department of Electrical Engineering and Computer Science, University of Michigan, Ann Arbor, MI, USA.

Biorxiv : the Preprint Server for Biology
|July 14, 2025
PubMed
Abstract

Insights

Freezing is a viable alternative to tissue fixation for myelin-sensitive quantitative MRI (qMRI). While qMRI values may slightly increase after thawing, histology confirms tissue integrity is maintained, supporting freezing for qMRI analysis.

Area of Science:

  • Neuroimaging
  • Histology
  • Biomedical Engineering

Background:

  • Quantitative MRI (qMRI) parameter validation typically uses fixed ex vivo tissue.
  • The impact of freezing and thawing on myelin-sensitive qMRI parameters and their histological correlation needs further investigation.

Purpose of the Study:

  • To evaluate the effects of freezing and thawing on myelin-sensitive qMRI parameters.
  • To compare qMRI results with histological staining (Luxol fast blue) in fresh, thawed, and fixed sheep brain tissue.

Main Methods:

  • Myelin water imaging, off-resonance RF saturation magnetization transfer (MT), and selective inversion recovery MT MRI were performed.
  • 14 sheep brain tissue samples were analyzed in fresh, thawed, and fixed states.
  • qMRI-derived myelin surrogate measures were compared to Luxol fast blue (LFB) staining.

Main Results:

  • qMRI parameters showed good correlation with LFB histology across all tissue states.
  • Thawed and fixed tissues displayed modest increases (3-32%) in most qMRI parameters compared to fresh tissue.
  • Histology confirmed that thawed samples retained tissue integrity after freezing.

Conclusions:

  • Freezing is a suitable alternative to fixation for myelin-sensitive qMRI analysis.
  • Freezing and thawing may cause differential changes in qMRI parameter values depending on regional myelin content.
  • Tissue integrity is preserved through freezing, making it a practical method for qMRI studies.
Keywords:
bi-exponential T1 mappingluxol fast blue histologymyelin water imaging (MWI)quantitative MRIquantitative magnetization transfer (qMT)tissue preparation

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