Creating synergy between established histology methods and newer spatial analysis modalities using fresh frozen...

Area of Science:

• Histology and Pathology
• Spatial Biology
• Mass Spectrometry Imaging

Background:

• Formalin-fixed paraffin-embedded (FFPE) tissues are standard but can hinder molecular analysis.
• Emerging spatial biology techniques increasingly require fresh-frozen samples.
• Label-free methods offer unique molecular insights but need optimized sample handling.

Purpose of the Study:

• To develop and validate a novel 3D printed cassette holder for cryostats.
• To enhance the preservation of fresh-frozen tissue integrity for spatial omics.
• To facilitate synergy between histology workflows and label-free spatial biology.

Main Methods:

• Development of 3D printed cassette holders for cryostat use.
• Embedding fresh-frozen tissue slices in hydroxypropyl methylcellulose polyvinylpyrrolidone.
• Testing the apparatus with fresh frozen tissue samples for subsequent analyses.

Main Results:

• The 3D printed cassette holders are compatible with commercially available cryostats.
• The protocol aids in preserving sample history throughout specimen analysis.
• The method helps curtail the decomposition of low-abundance endogenous chemical moieties.

Conclusions:

• The developed 3D printed cassette holder protocol optimizes fresh-frozen tissue handling for spatial omics.
• This approach supports the preservation of critical molecular information for mass spectrometry-based analyses.
• The protocol offers a valuable tool for biomedical discovery programs integrating histology and advanced spatial biology.