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Open Access Highly Accessed Research article

Preservation of biomolecules in breast cancer tissue by a formalin-free histology system

Mehdi Nassiri1*, Sharon Ramos1, Hajir Zohourian1, Vladimir Vincek2, Azorides R Morales1 and Mehrdad Nadji1

Author affiliations

1 Department of Pathology, University of Miami Miller School of Medicine, Miami, Florida, USA

2 Department of Pathology, University of Florida, Gainesville, Florida, USA

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Citation and License

BMC Clinical Pathology 2008, 8:1  doi:10.1186/1472-6890-8-1

Published: 29 January 2008



The potential problems associated with the use of formalin in histology, such as health hazards, degradation of RNA and cross-linking of proteins are well recognized. We describe the utilization of a formalin-free fixation and processing system for tissue detection of two important biopredictors in breast cancer – estrogen receptor and HER2 – at the RNA and protein levels.


Parallel sections of 62 cases of breast cancer were fixed in an alcohol-based molecular fixative and in formalin. Molecular fixative samples were processed by a novel formalin-free microwave-assisted processing system that preserves DNA, RNA and proteins. Formalin-fixed samples were processed using the conventional method. Estrogen receptor was assessed by immunohistochemistry and real-time PCR. HER2 was assessed by immunohistochemistry, FISH, CISH and real-time PCR.


The immunohistochemical reaction for estrogen receptor was similar in molecular- and formalin-fixed samples (Spearman Rank R = 0.83, p < 0.05). Also HER2 result was similar to that of formalin-fixed counterparts after elimination of antigen retrieval step (Spearman Rank R = 0.84, p < 0.05). The result of HER2 amplification by FISH and CISH was identical in the molecular fixative and formalin-fixed samples; although a shorter digestion step was required when using the former fixative. Real-time PCR for both estrogen receptor and HER2 were successful in all of the molecular fixative specimens.


The formalin-free tissue fixation and processing system is a practical platform for evaluation of biomolecular markers in breast cancer and it allows reliable DNA and RNA and protein studies.