Berrino E.a,b· Annaratone L.a,b· Detillo P.c· Grassini D.a,b· Bragoni A.a,b· Sapino A.a,b· Bussolati B.d· Bussolati G.b· Marchiò C.a,b

Author affiliations

aCandiolo Cancer Institute, FPO-IRCCS, Candiolo, Italy
bDepartment of Medical Sciences, University of Turin, Turin, Italy
cADDAX Biosciences srl., Turin, Italy
dDepartment of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy

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Article / Publication Details

First-Page Preview

Received: April 11, 2022
Accepted: June 10, 2022
Published online: July 20, 2022

Number of Print Pages: 11
Number of Figures: 4
Number of Tables: 0

ISSN: 1015-2008 (Print)
eISSN: 1423-0291 (Online)

For additional information: https://www.karger.com/PAT

Abstract

Introduction: Optimization of pre-analytic procedures and tissue processing is a basic requirement for reliable and reproducible data to be obtained. Tissue fixation in formalin represents the extensively favored method for surgical tissue specimen processing in diagnostic pathology; however, formalin fixation exerts a blasting effect on DNA and RNA. Methods: A formic acid-deprived formaldehyde solution was prepared by removing acids with an ion-exchange basic resin and the concentrated, acid-deprived formaldehyde (ADF) solution was employed to prepare a 4% ADF solution in 0.1 M phosphate buffer, pH 7.2–7.4. Human (n = 27) and mouse (n = 20) tissues were fixed in parallel and similar conditions in either ADF or neutral buffered formalin (NBF). DNAs and RNAs were extracted, and fragmentation analyses were performed. Results: Besides no significant differences in terms of extraction yield and absorbance ratio, ADF fixation reduced DNA fragmentation, i.e., the largest fragments (>5,000 bp) were significantly more prevalent in the DNAs purified from ADF-fixed tissues (p < 0.001 in both cohorts). Moreover, we observed that DNA preservation is more stable in ADF-fixed tissue compared to NBF-fixed tissues. Conclusion: Although DNA fragmentation in FFPE tissues is a multifactor process, we showed that the removal of formic acid is responsible for a significant improvement in DNA preservation.

© 2022 S. Karger AG, Basel

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First-Page Preview

Received: April 11, 2022
Accepted: June 10, 2022
Published online: July 20, 2022

Number of Print Pages: 11
Number of Figures: 4
Number of Tables: 0

ISSN: 1015-2008 (Print)
eISSN: 1423-0291 (Online)

For additional information: https://www.karger.com/PAT

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