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Evaluation of non-formalin tissue fixation for molecular profiling studies
John W Gillespie1, Carolyn J M Best, Verena E Bichsel
1Pathogenetics Unit, Laboratory of Pathology, National Cancer Institute, Rm. 2C500. Bldg. 10, 9000 Rockville Pike, Bethesda, MD 20892, USA.
The American Journal of Pathology
|February 13, 2002
Summary
70% ethanol fixation and paraffin embedding is effective for molecular profiling. This method allows for the recovery of DNA, mRNA, and proteins from clinical tissue specimens for downstream analyses.
Area of Science:
- Biotechnology
- Molecular Biology
- Histology
Background:
- Clinical tissue specimens are crucial for molecular profiling.
- Standard fixation methods may impact molecular analysis.
- Evaluating alternative fixation techniques is necessary.
Purpose of the Study:
- To assess the efficacy of 70% ethanol fixation and paraffin embedding for molecular profiling.
- To compare the molecular yield from ethanol-fixed tissues with snap-frozen and formalin-fixed specimens.
Main Methods:
- Proteomic analysis using gel electrophoresis and immunoblotting.
- Nucleic acid recovery and analysis via PCR.
- Histological evaluation of fixed tissues.
Main Results:
- Ethanol-fixed, paraffin-embedded tissues yielded comparable protein profiles to snap-frozen samples, with slightly reduced quantity.
- mRNA recovery was reduced but sufficient for reverse transcription PCR, outperforming formalin fixation.
- DNA recovery was superior in ethanol-fixed specimens compared to formalin-fixed ones.
Conclusions:
- 70% ethanol fixation and paraffin embedding is a viable method for molecular profiling.
- This technique enables the recovery of sufficient DNA, mRNA, and proteins for various downstream analyses.
- The method provides good histology and is a valuable alternative for tissue specimen preservation.