Impact of standardization in tissue processing: the performance of different fixatives

Most tissues in clinical practice are formalin-fixed and paraffin-embedded for histological as well as molecular analyses. The reproducibility and uniformity of molecular analyses is strictly dependent on the quality of the biomolecules, which is highly influenced by pre-analytical processes. In thi...

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Bibliographic Details
Published in:New biotechnology 2022-11, Vol.71, p.30-36
Main Authors: De Martino, Eleonora, Medeot, Caterina, D’Amico, Lorenzo, Stanta, Giorgio, Bonin, Serena
Format: Article
Language:English
Subjects:
Bouin
Formalin
ISO standard
Paraffin embedding
Pre-analytical processes
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Summary:Most tissues in clinical practice are formalin-fixed and paraffin-embedded for histological as well as molecular analyses. The reproducibility and uniformity of molecular analyses is strictly dependent on the quality of the biomolecules, which is highly influenced by pre-analytical processes. In this study, the effect of different fixatives was compared, including formalin, Bouin’s solution, RCL2® and TAG-1™ fixatives, by stringent application of ISO standards in mouse liver tissue processing, including formalin-free transport of tissues and tissue grossing in a refrigerated environment. The effect of fixatives was studied in terms of nucleic acid quality at the time of tissue processing and after one year of tissue storage at room temperature in the dark. Furthermore, a microcomputed tomography (CT) scan analysis was applied to investigate the paraffin embedding. The results show that the application of ISO standards in tissue processing allows analysis of 400 bases amplicons from RNA and 1000 bases from DNA, even in extracts from formalin-fixed and paraffin-embedded tissues. However, after one year storage at room temperature in the dark, a degradation of the nucleic acids was observed. Nevertheless, extracts can still be analyzed, but for metachronous tests it is highly recommended to repeat the quantitation of housekeeping genes in order to standardize the extent of nucleic acid degradation. •ISO standards application for pre-analytical conditions allows analyzing 400 bases RNA stretches from FFPE tissues.•ISO standards application for pre-analytical conditions allows analyzing 1000 bases DNA stretches from FFPE tissues.
ISSN:1871-6784
1876-4347
DOI:10.1016/j.nbt.2022.07.001