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Formaldehyde substitute fixatives. Analysis of macroscopy, morphologic analysis, and immunohistochemical analysis
Cathy B Moelans1, Natalie ter Hoeve, Jan-Willem van Ginkel
1Dept of Pathology (H04.131), University Medical Center Utrecht, Heidelberglaan 100, Utrecht, the Netherlands.
American Journal of Clinical Pathology
|September 16, 2011
Summary
Three new alcohol-based fixatives were evaluated against neutral buffered formalin (NBF) for tissue staining. None fully matched NBF, with each showing unique drawbacks for histology and immunohistochemistry.
Area of Science:
- Histology and Pathology
- Biochemical and Molecular Biology
Background:
- Formaldehyde fixation, the standard, poses toxicity and cross-linking issues impacting immunohistochemical studies.
- Alternative alcohol-based fixatives are sought to mitigate formaldehyde's drawbacks.
Purpose of the Study:
- To compare the efficacy of three novel alcohol-based fixatives (F-Solv, FineFIX, RCL2) against neutral buffered formalin (NBF) for routine tissue staining.
- To evaluate their impact on histochemical and immunohistochemical staining.
Main Methods:
- Fresh tissue samples were fixed using F-Solv, FineFIX, RCL2, and NBF for varying durations.
- Standard histological procedures including Hematoxylin and Eosin (H&E) staining were performed.
- Seven histochemical and nine immunohistochemical stains were applied to assess antigenicity and morphology.
Main Results:
- RCL2 fixation yielded soft tissues, complicating sectioning.
- F-Solv and FineFIX caused partial tissue disintegration.
- F-Solv required significant protocol adjustments for comparable morphology to NBF.
- FineFIX induced shrinkage, degranulation, and red blood cell (RBC) lysis.
- RCL2 showed good morphology but caused pigment deposition and RBC lysis.
Conclusions:
- None of the tested alcohol-based fixatives (F-Solv, FineFIX, RCL2) were universally superior to NBF for routine staining and immunohistochemistry.
- Each alternative fixative presented specific limitations, including tissue handling, morphological artifacts, and compatibility issues.
- Further optimization and consideration of environmental factors are necessary for potential adoption of these fixatives.
