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Gene Expression and Cancer Classification · Journal article
PLOS One · August 14, 2026
Encouraging direction, but not yet definitive.
This study demonstrates that supercritical CO₂-based formalin-free tissue embedding produces higher DNA integrity (measured by DIN values) than conventional formalin-fixed paraffin-embedded tissue, and maintains RNA suitable for next-generation sequencing across multiple timepoints. However, RNA quality by RIN values was lower in shorter timeframes, and clinical utility for cancer diagnosis and treatment guidance remains undemonstrated.
Laboratory methods comparison study with temporal analysis. Tissue samples; specific tissue types, histology, clinical sources, and number of specimens not stated.. Intervention: Supercritical CO₂-based tissue embedding in a xylene and formalin-free manner. Compared with: Conventional formalin-fixed paraffin-embedded tissues and non-fixed tissues.
DNA integrity (DIN values) was higher in formalin-free embedded tissues than in conventionally processed tissues RNA quality (RIN values) was lower in 3-day and 2-year-old non-fixed tissues compared to formalin-fixed tissues but similar in 7-year-old tissues RNA isolated from 3-day, 2-year, and most 7-year-old non-formalin-fixed tissues was suitable for next-generation sequencing
Safety was not reported in the material analysed. Check the source before drawing any conclusion about harm.
If validated prospectively, this method could improve molecular diagnostics in pathology by providing better DNA and adequate RNA integrity without formaldehyde exposure. Current evidence does not yet support clinical decision-making, as diagnostic accuracy or therapeutic impact has not been assessed.
A sound experimental comparison of a novel tissue-embedding technique showing superior DNA integrity and adequate RNA quality for molecular diagnostics, but limited by single-centre design, lack of clinical outcome data, and incomplete characterization of long-term stability.
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Quoted from the source exactly as published.
If validated prospectively, this method could improve molecular diagnostics in pathology by providing better DNA and adequate RNA integrity without formaldehyde exposure. Current evidence does not yet support clinical decision-making, as diagnostic accuracy or therapeutic impact has not been assessed.
Graded across the dimensions that decide whether you should act, each from what the source actually supports. There is no single score, and where a dimension was not assessed it says so.
In recent decades, the field of pathology has been enriched by the development of molecular biology, enabling increased specificity in cancer diagnosis and personalized therapy based on the molecular profile of tumor cells. Traditional tissue embedding techniques, routinely using toxic and even carcinogenic substances, in particular formaldehyde, are still standard practice for the field of pathology and consequently for molecular pathology. Formaldehyde damages DNA and RNA, resulting in suboptimal molecular diagnostics, possibly impacting cancer diagnosis and treatment quality. A novel tissue embedding technique based on supercritical CO 2 can be used in a formaldehyde-free manner, producing high-quality samples suitable for basic pathology techniques. Early data indicates that the DNA isolated from tissues processed in this manner can be used for molecular pathology, but detailed DNA analysis lacked, and RNA had not been analyzed at all. Here we performed integrity analysis of both the DNA and RNA isolated from tissues 7 years, 2 years, and 3 days after embedding and compared non-fixed and formalin-fixed tissues. The results show that this novel embedding technique, used in xylene and formalin-free manner, results in DNA of higher quality than that of conventionally processed tissues, as shown by higher DIN values. On the other hand, RNA quality, as shown by RIN values, was generally lower in 3-day and 2-year old, non-fixed (NFPE) tissues compared to formalin-fixed (FFPE) tissues but similar in 7 year-old tissues. RNA isolated from 3-day, 2-year, and even most 7-year-old non-formalin fixed tissues was still suitable for the main downstream molecular technique Next-Generation-Sequencing.
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