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

Authors
 Joon-Yong Chung  ;  Joon Seon Song  ;  Kris Ylaya  ;  John D. Sears  ;  Lauren Choi  ;  Hanbyoul Cho  ;  Avi Z. Rosenberg  ;  Stephen M. Hewitt* 
Citation
 JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, Vol.66(2) : 121-135, 2018 
Journal Title
 JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY 
ISSN
 0022-1554 
Issue Date
2018
Keywords
RNA integrity ; alcohol ; fixation ; formaldehyde ; histology ; immunohistochemistry ; protein
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.
Full Text
https://journals.sagepub.com/doi/full/10.1369/0022155417741467
DOI
10.1369/0022155417741467
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Obstetrics and Gynecology (산부인과학교실) > 1. Journal Papers
Yonsei Authors
Cho, Hanbyoul(조한별) ORCID logo https://orcid.org/0000-0002-6177-1648
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/165337
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