Effect of Fixation to the Degradation of Nuclear and Mitochondrial DNA in Different Tissues.

Author(s): Miething F, Hering S, Hanschke B, Dressler J

Publication: J Histochem Cytochem, 2006, Vol. 54, Page 371

PubMed ID: 16260588 PubMed Review Paper? No

Purpose of Paper

Comparative analysis of DNA and cellular morphology preservation in different tissues fixed with seven different fixatives for various durations (1 to 336 days).

Conclusion of Paper

While Carnoy's solution and glutaraldehyde were superior in the preservation of DNA (nuclear and mitochondrial, mt), cellular structure was compromised. Fixation with 4% phosphate buffered formalin yielded intermediate DNA preservation and superior cellular structure. In terms of tissue type, skeletal muscle and kidney yielded the most reliable results possibly due to tissue-dependent variations in fixative penetration.

Studies

Study Purpose

Comparative analysis of DNA preservation (using the FGA, TC11, and HV1 PCR platforms) after fixation in seven different fixatives (including buffered and unbuffered formalin, Carnoy's solution, and glutaraldehyde) for various durations (1-336 days).

Summary of Findings:

Carnoy's solution and glutaraldehyde were superior to other fixatives in the preservation of DNA integrity, with amplification of mt (220 bp) and nuclear (800 bp) products possible after 336 days of fixation for all tissues investigated. After fixation with 4% phosphate buffered formalin for 336 days, mtDNA remained detectable, although nuclear DNA was not detectable beyond 28 days of fixation.

Biospecimens

Preservative Types

Formalin
Other Preservative

Diagnoses:

Autopsy
Normal

Platform:

Analyte	Technology Platform
DNA	PCR

Pre-analytical Factors:

Classification	Pre-analytical Factor	Value(s)
Biospecimen Preservation	Time in fixative	1 day 3 days 5 days 7 days 14 days 28 days 56 days 84 days 168 days 336 days
Biospecimen Preservation	Type of fixation/preservation	4% phosphate buffered formalin 4% unbuffered formalin Alcohol-formic acid solution Potassium dichromate acetic acid Davidson's solution Carnoy's solution Glutaraldehyde

Study Purpose

Comparative analysis of cellular structure after fixation in seven different fixatives (including buffered and unbuffered formalin, Carnoy's solution, and glutaraldehyde) for various durations (1-336 days).

Summary of Findings:

4% phosphate buffered formalin was superior in the preservation of cellular structure compared to other fixatives examined. Cellular structure was poorly preserved with Carnoy's solution and glutaraldehyde.

Biospecimens

Preservative Types

Formalin
Other Preservative

Diagnoses:

Autopsy
Normal

Platform:

Analyte	Technology Platform
Morphology	H-and-E microscopy

Pre-analytical Factors:

Classification	Pre-analytical Factor	Value(s)
Biospecimen Preservation	Type of fixation/preservation	Davidson's solution Carnoy's solution Glutaraldehyde 4% phosphate buffered formalin 4% unbuffered formalin Alcohol-formic acid solution Potassium dichromate acetic acid

Study Purpose

Comparative analysis of DNA preservation and cellluar structure in five different tissue types (cardiac muscle, skeletal muscle, liver, kidney, brain) after fixation in seven different fixatives for various durations.

Summary of Findings:

Skeletal muscle and kidney yielded the most reliable preservation of DNA and cellular structure across the fixatives examined.

Biospecimens

Preservative Types

Formalin

Diagnoses:

Normal
Autopsy

Platform:

Analyte	Technology Platform
DNA	PCR
Morphology	H-and-E microscopy

Pre-analytical Factors:

Classification	Pre-analytical Factor	Value(s)

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