Effect of Formalin, Acetone, and RNAlater Fixatives on Tissue Preservation and Different Size Amplicons by Real-Time PCR from Paraffin-Embedded Tissue.

Author(s): Paska C, Bogi K, Szilak L, Tokes A, Szabo E, Sziller I, Rigo J, Sobel G, Szabo I, Kaposi-Novak P, Kiss A, Schaff Z

Publication: Diagn Mol Pathol, 2004, Vol. 13, Page 234

PubMed ID: 15538114 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   Comparative analysis of the preservation of cellular structure among paraffin embedded tissues fixed by different methods (formalin, acetone, RNAlater).

   Summary of Findings:

   Structural integrity was adequately preserved by all fixatives examined, although the degree of nuclear and cytoplasmic detail was superior with formalin fixation.

   Biospecimens

   • Tissue - Endometrium

   Preservative Types

   • Formalin
   • Other Preservative
   • RNAlater

   Diagnoses:

   • Neoplastic - Benign

   Platform:

   Analyte	Technology Platform
   Morphology	H-and-E microscopy

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Formalin (buffered) RNAlater Acetone

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2. Study Purpose

   Comparative analysis of immunohistochemical staining (claudin 4 and 7) among paraffin embedded tissues fixed by various methods (formalin, RNAlater, acetone).

   Summary of Findings:

   Immunoreactivity (claudin 4 and 7) was superiorly preserved with formalin fixation, which yielded more robust and defined signals. Fixation in RNAlater resulted in uneven and diffuse staining in an antibody dependent manner.

   Biospecimens

   • Tissue - Endometrium

   Preservative Types

   • Formalin
   • Other Preservative
   • RNAlater

   Diagnoses:

   • Neoplastic - Benign

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Formalin (buffered) RNAlater Acetone

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3. Study Purpose

   Comparative analysis of gene expression levels (GADPH, beta-globin) in paraffin embedded tissues fixed by various methods (formalin, RNAlater, acetone). The influence of amplicon size on detectable cDNA levels was also addressed.

   Summary of Findings:

   Fixative type did not impact detectable levels of gene expression. Gene expression levels were inversely proportional to amplicon size, ie. shorter amplicons were more abundant. Further, the longest amplicon (536 bp) was not detected in the majority of samples examined.

   Biospecimens

   • Tissue - Endometrium

   Preservative Types

   • Formalin
   • RNAlater
   • Other Preservative

   Diagnoses:

   • Neoplastic - Benign

   Platform:

   Analyte	Technology Platform
   RNA	Real-time qRT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Formalin (buffered) RNAlater Acetone
   Real-time qRT-PCR Specific	Targeted nucleic acid	GAPDH beta-globin
   Real-time qRT-PCR Specific	Length of gene fragment	121 bp 166 bp 225 bp 310 bp 406 bp 536 bp

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