Formaldehyde Substitute Fixatives: Analysis of Macroscopy, Morphologic Analysis, and Immunohistochemical Analysis

Because formaldehyde is toxic and creates cross-links that may hinder immunohistochemical studies, we tested 3 new cross-linking (F-Solv [Adamas, Rhenen, the Netherlands]) and non-cross-linking (FineFIX [Milestone, Bergamo, Italy] and RCL2 [Alphelys, Plaisir, France]) alcohol-based fixatives for rou...

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Bibliographic Details
Published in:	American journal of clinical pathology 2011-10, Vol.136 (4), p.548-556
Main Authors:	MOELANS, Cathy B, TER HOEVE, Natalie, VAN GINKEL, Jan-Willem, TEN KATE, FieboJ, VAN DIEST, Paul J
Format:	Article
Language:	eng
Subjects:	Biological and medical sciences Fixatives Formaldehyde Humans Immunohistochemistry Investigative techniques, diagnostic techniques (general aspects) Medical sciences Paraffin Embedding Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques Tissue Fixation - methods
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Summary:	Because formaldehyde is toxic and creates cross-links that may hinder immunohistochemical studies, we tested 3 new cross-linking (F-Solv [Adamas, Rhenen, the Netherlands]) and non-cross-linking (FineFIX [Milestone, Bergamo, Italy] and RCL2 [Alphelys, Plaisir, France]) alcohol-based fixatives for routine staining in comparison with neutral buffered formalin (NBF) as the "gold standard." Fresh tissue samples were divided into 4 equal pieces and fixed in all fixatives for varying times. After paraffin embedding, H&E staining, 7 common histochemical stains, and 9 common immunohistochemical stains were performed. RCL2 fixation resulted in soft and slippery tissue, causing sectioning difficulties. F-Solv and FineFIX led to partial tissue disintegration during fixation. F-Solv performed morphologically similar to NBF but needed considerable protocol adjustments before being applicable in daily histologic and immunohistochemical practice. FineFIX did not necessitate major protocol changes but caused shrinkage artifacts, degranulation, and lysis of RBCs. RCL2 generated morphologically overall good results without major protocol changes but caused pigment deposition, degranulation, and RBC lysis. The alcohol-based fixatives had positive and negative attributes and environmental drawbacks, and none was overall comparable to NBF with regard to macroscopy, morphologic evaluation, and immunohistochemical studies.
ISSN:	0002-9173 1943-7722