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Ethanol fixation method for heart and lung imaging in micro-CT

Matej Patzelt1,2,3, Jana Mrzilkova4,5,6, Jan Dudak1,2,7,8

  • 1Specialized Laboratory of Experimental Imaging, Third Faculty of Medicine, Charles University, Prague, Czech Republic.

Japanese Journal of Radiology
|March 16, 2019
PubMed
Abstract

Insights

Simple ethanol fixation enhances soft tissue imaging in micro-computed tomography (micro-CT) without contrast agents. Ascending ethanol concentrations provide the best detailed visualization of organs for micro-CT analysis.

Area of Science:

  • Biomedical Imaging
  • Radiology
  • Histology

Background:

  • Micro-computed tomography (micro-CT) imaging of soft tissues is limited by low intrinsic contrast.
  • Enhancing soft tissue visibility in micro-CT typically requires contrast agents, which can be complex or unavailable for ex vivo studies.

Purpose of the Study:

  • To develop a straightforward staining method for ex vivo soft tissue imaging using micro-CT.
  • To achieve improved contrast without relying on traditional contrast agents.

Main Methods:

  • Mouse hearts and lungs (n=30) were utilized for the study.
  • Organs underwent fixation in 97% ethanol, 50% ethanol, or a series of ascending ethanol concentrations.
  • Micro-CT imaging was performed at various time points (72, 168, 336 hours) and compared to native samples.

Main Results:

  • Ethanol fixation consistently enhanced contrast across all tested methods.
  • Fixation in 97% ethanol provided rapid contrast but led to significant tissue hardening.
  • Fixation in 50% ethanol showed delayed contrast enhancement and poorer detail visualization.
  • A graded series of ascending ethanol concentrations yielded optimal results, revealing intricate details without tissue damage.

Conclusions:

  • Simple ethanol fixation is an effective method for improving soft tissue contrast in micro-CT.
  • Fixing soft tissue samples in a series of ascending ethanol concentrations provides the most favorable outcomes for detailed imaging.
Keywords:
EthanolHeartLungsMiceMicro-CT

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