title: Weigners fixative-an alternative to formalin fixation for histology with improved preservation of nucleic acids.
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.
authors.affiliation: Department of Veterinary Pathology, Freie Universität Berlin, Berlin, Germany. robert.klopfleisch@fu-berlin.de
authors.forename: R
authors.initials: R
authors.lastname: Klopfleisch
authors.forename: M
authors.initials: M
authors.lastname: von Deetzen
authors.forename: A Th
authors.initials: AT
authors.lastname: Weiss
authors.forename: J
authors.initials: J
authors.lastname: Weigner
authors.forename: F
authors.initials: F
authors.lastname: Weigner
authors.forename: J
authors.initials: J
authors.lastname: Plendl
authors.forename: A D
authors.initials: AD
authors.lastname: Gruber
mesh_terms: D000818:Animals; D002415:Cats; D004247:DNA / Q000302:isolation & purification; D004285:Dogs; D005404:Fixatives; D005557:Formaldehyde; D007150:Immunohistochemistry; D007668:Kidney / Q000737:chemistry; D035683:MicroRNAs / Q000302:isolation & purification; D009696:Nucleic Acids / Q000302:isolation & purification* / Q000592:standards; D016612:Paraffin Embedding; D011309:Preservation, Biological / Q000379:methods* / Q000592:standards; D012333:RNA, Messenger / Q000302:isolation & purification; D016707:Tissue Fixation / Q000379:methods*
chemical_list: D005404:Fixatives; D035683:MicroRNAs; D009696:Nucleic Acids; D012333:RNA, Messenger; D005557:Formaldehyde; D004247:DNA