Cervical Tissue Collection Methods for RNA Preservation: Comparison of Snap-frozen, Ethanol-fixed, and RNAlater fixation.

Author(s): Wang S, Sherman ME, Rader J, Carreon J, Schiffman M, Baker C

Publication: Diagn Mol Path, 2006, Vol. 15, Page 144

PubMed ID: 16932069 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to assess RNA quality in benign and cancerous cervical tissue that were (1) snap frozen, OCT-embedded, and stored at -80 degrees C, (2) fixed in ethanol, embedded in low melt polyester wax, and stored at 4 degrees C or (3) preserved in RNAlater and stored at -20 degrees C.

   Summary of Findings:

   The authors conclude that cervix specimens fixed in RNAlater and those snap frozen in OCT produced RNA of comparable quality; however, it is worthy to note that while both 28S and 18S rRNA bands were present in differentially cryopreserved specimens 28S/18S rRNA ratios differed, 1.64 versus 0.79 in liquid nitrogen and RNAlater preserved specimens respectively. Conversely, RNA isolated from ethanol fixed tissue was degraded, and a distinct 28S ribosomal RNA band was not observed.

   Biospecimens

   • Tissue - Cervix

   Preservative Types

   • OCT
   • RNAlater
   • Ethanol

   Diagnoses:

   • Neoplastic - Carcinoma
   • Neoplastic - Benign

   Platform:

   Analyte	Technology Platform
   RNA	Automated electrophoresis/Bioanalyzer

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Embedding medium	OCT Low melt polyester wax
   Biospecimen Preservation	Type of fixation/preservation	Ethanol RNAlater OCT
   Storage	Storage temperature	4 degrees C -20 degrees C -80 degrees C

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2. Study Purpose

   To assess the preservation of cellular structure and the technical constraints of embedding in OCT versus wax.

   Summary of Findings:

   Snap frozen OCT-embedded tissue was the easiest and least time intensive to process due to its positioning and orientation at the time of collection, and the preservation of cellular structure was adequate. Orientation of both ethanol and RNAlater fixed tissue was difficult. While ethanol fixed tissue yielded superior preservation of cellular structure, wax embedding and sectioning was time intensive. Cellular structure was adequately preserved in RNAlater fixed specimens.

   Biospecimens

   • Tissue - Cervix

   Preservative Types

   • OCT
   • RNAlater
   • Ethanol

   Diagnoses:

   • Neoplastic - Carcinoma
   • Neoplastic - Benign

   Platform:

   Analyte	Technology Platform
   Morphology	H-and-E microscopy

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Embedding medium	OCT Low melt polyester wax
   Biospecimen Preservation	Type of fixation/preservation	Snap frozen Ethanol RNAlater

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