Histomorphological and Molecular Assessments of the Fixation Times Comparing Formalin and Ethanol-Based Fixatives

Overview

Journal J Histochem Cytochem

Publisher Sage Publications

Specialty Biochemistry

Date 2017 Nov 11

PMID 29125916

Citations 15

Authors

Joon-Yong Chung

Joon Seon Song

Kris Ylaya

John D Sears

Lauren Choi

Hanbyoul Cho

Avi Z Rosenberg

Stephen M Hewitt

Affiliations

Soon will be listed here.

Abstract

The lack of standardization of tissue handling and processing hinders the development and validation of new biomarkers in research and clinical settings. We compared the histomorphology and the quality and quantity of biomolecules in paraffin-embedded mouse tissues, followed by fixation with neutral buffered formalin (NBF), 70% ethanol, and buffered ethanol (BE70) fixative. The quality of the histomorphology and immunohistochemistry in BE70 was relatively time-independent, whereas those in NBF rapidly decreased after 1 week of fixation. Protein recovered from tissue fixed in 70% ethanol and BE70 was compatible with Western blot and protein array using AKT and GAPDH antibodies, regardless of the fixation time. In addition, the quality and quantity of RNA extracted from tissue in ethanol-based fixative showed minimal changes from 4 hr to 6 months, whereas NBF had a dramatic detrimental change in RNA quality after 1 week of fixation. Furthermore, ethanol-based fixative offers a superior DNA template for PCR amplification-based molecular assays than NBF. In conclusion, coagulative, ethanol-based fixatives show a broader time spectrum than the aldehyde crosslinking fixative NBF in their histomorphological features and the quantity and quality of the biomolecules from paraffin-embedded tissue, and they may facilitate the use of fixative-fixed paraffin-embedded tissues in research and clinical laboratories, avoiding overfixation.

Citing Articles

Effects of fixation and demineralization on histomorphology and DNA amplification of canine bone marrow.

Diamantino G, Beeler-Marfisi J, Foster R, Sears W, Defarges A, Vernau W Vet Pathol. 2024; 61(6):943-951.

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Evaluation of a Novel Fixative Solution for Liquid-Based Cytology in Diagnostic Cytopathology.

Casatta N, Poli A, Bassani S, Veronesi G, Rossi G, Ferrari C Diagnostics (Basel). 2023; 13(24).

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Infrared spectroscopic laser scanning confocal microscopy for whole-slide chemical imaging.

Yeh K, Sharma I, Falahkheirkhah K, Confer M, Orr A, Liu Y Nat Commun. 2023; 14(1):5215.

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Gelatin Zymography Can Be Performed on Fixed Brain Tissue.

Chen Z, Chuang D, Chen S, He Q, Tomlison B, Cui J J Histochem Cytochem. 2023; 71(9):481-493.

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The Application of Guanidinium to Improve Biomolecule Quality in Fixed, Paraffin-embedded Tissue.

Chung J, Kim K, Ylaya K, Walker-Bawa K, Perry C, Star R J Histochem Cytochem. 2023; 71(2):87-101.

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