Rapid two-temperature formalin fixation

Formalin fixation is a mainstay of modern histopathologic analysis, yet the practice is poorly standardized and a significant potential source of preanalytical errors. Concerns of workflow and turnaround time drive interest in developing shorter fixation protocols, but rapid protocols can lead to po...

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Bibliographic Details
Published in:PloS one 2013-01, Vol.8 (1), p.e54138-e54138
Main Authors: Chafin, David, Theiss, Abbey, Roberts, Esteban, Borlee, Grace, Otter, Michael, Baird, Geoffrey S
Format: Article
Language:English
Subjects:
Analysis
Antigens
Assaying
Biochemistry
Biology
Breast cancer
Chemical reactions
Chemistry
Downstream effects
Enzymes
Epitopes
Fixation
Fixatives
Formaldehyde
Heat
Histopathology
Historical account
Humans
Immunohistochemistry
Immunohistochemistry - methods
Incubation
Laboratories
Medicine
Palatine Tonsil - metabolism
Proteins
Proto-Oncogene Proteins c-bcl-2 - analysis
Protocol
Reproducibility of Results
Temperature
Time Factors
Tissue Fixation - methods
Variables
Workflow
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Summary:Formalin fixation is a mainstay of modern histopathologic analysis, yet the practice is poorly standardized and a significant potential source of preanalytical errors. Concerns of workflow and turnaround time drive interest in developing shorter fixation protocols, but rapid protocols can lead to poor histomorphology or inadequate downstream assay results. Additionally, assays such as immunohistochemistry for phosphorylated epitopes have historically been challenging in the context of formalin-fixed tissue, indicating that there may be room for improvement in this process that is fundamental to the practice of anatomic pathology. With these issues in mind, we studied basic formalin biochemistry to develop a novel formalin fixation protocol that involves a pre-incubation in subambient temperature formalin prior to a brief exposure to heated formalin. This new protocol is more rapid than standard protocols yet preserves histomorphology and yields tissue that is compatible with an expanded set of downstream clinical and research assays, including immunohistochemistry for phosphorylated epitopes.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0054138