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Updated: May 22, 2026

Protocol for HER2 FISH Using a Non-cross-linking, Formalin-free Tissue Fixative to Combine Advantages of Cryo-preservation and Formalin Fixation
Published on: December 25, 2017
Weigners fixative-an alternative to formalin fixation for histology with improved preservation of nucleic acids
R Klopfleisch1, M von Deetzen, A Th Weiss
1Department of Veterinary Pathology, Freie Universität Berlin, Berlin, Germany. robert.klopfleisch@fu-berlin.de
Abstract:
Formalin fixation and paraffin embedding (FFPE) is the standard method for tissue storage in histopathology. However, FFPE has disadvantages in terms of user health, environment, and nucleic acid integrity. Weigners fixative has been suggested as an alternative for embalming cadavers in human and veterinary anatomy. The present study tested the applicability of Weigners for histology and immunohistochemistry and the preservation of nucleic acids. To this end, a set of organs was fixed for 2 days and up to 6 months in Weigners (WFPE) or formalin. WFPE tissues from the skin, brain, lymphatic tissues, liver, and muscle had good morphologic preservation, comparable to formalin fixation. The quality of kidney and lung samples was inferior to FFPE material due to less accentuated nuclear staining and retention of proteinaceous interstitial fluids. Azan, Turnbull blue, toluidin, and immunohistochemical stainings for CD79a, cytokeratin, vimentin, and von Willebrand factor led to comparable results with both fixates. Of note, immunohistochemical detection of CD3 was possible after 6 months in WFPE but not in FFPE tissues. mRNA, miRNA, and DNA from WFPE tissues had superior quality and allowed for amplification of miRNA, 400-bp-long mRNA, and 1000-bp-long DNA fragments after 6 months of fixation in WFPE. In summary, Weigners fixative is a nonhazardous alternative to formalin, which provides a good morphologic preservation of most organs, a similar sensitivity for protein detection, and a superior preservation of nucleic acids. Weigners may therefore be a promising alternative to cryopreservation and may be embraced by people affected by formalin allergies.
Insights
Weigners fixative offers a safer, non-hazardous alternative to formalin for tissue preservation. This method provides good morphology and superior nucleic acid preservation, making it suitable for histology and immunohistochemistry.
Area of Science:
- Histopathology and Molecular Biology
Background:
- Formalin fixation and paraffin embedding (FFPE) is standard but has health, environmental, and nucleic acid integrity drawbacks.
- Weigners fixative is a potential alternative, previously used in anatomy, with its applicability for histology needing evaluation.
Purpose of the Study:
- To assess Weigners fixative's suitability for histology and immunohistochemistry.
- To evaluate nucleic acid preservation in tissues fixed with Weigners fixative.
Main Methods:
- Organs were fixed in Weigners fixative (WFPE) or formalin for up to 6 months.
- Morphological preservation was assessed.
- Histological and immunohistochemical stainings were performed.
- Nucleic acids (mRNA, miRNA, DNA) were extracted and analyzed for quality and amplifiability.
Main Results:
- WFPE showed good morphological preservation in skin, brain, lymphatic tissues, liver, and muscle, comparable to FFPE.
- Kidney and lung samples had inferior quality with WFPE.
- Histology and immunohistochemistry (CD79a, cytokeratin, vimentin, vWF) yielded comparable results for both fixatives.
- Immunohistochemical detection of CD3 was successful after 6 months in WFPE, but not FFPE.
- WFPE tissues demonstrated superior nucleic acid quality, enabling amplification of miRNA, 400-bp mRNA, and 1000-bp DNA fragments after 6 months.
Conclusions:
- Weigners fixative is a non-hazardous alternative to formalin for tissue fixation.
- It offers good morphological preservation for most organs and comparable protein detection sensitivity.
- WFPE provides superior nucleic acid preservation, making it a promising alternative to cryopreservation, especially for those with formalin allergies.

