Abstract
Molecular analysis of tissue lesions is increasingly used in laboratories to identify new prognostic and therapeutic markers. Formalin has long been the tissue fixative of choice in the laboratories of pathology, as it preserves tissue morphology allowing accurate histological diagnosis. However, formalin is highly toxic and alters nucleic acids and protein integrity, so that new fixatives are critically needed that would allow both morphological and molecular analysis on the same tissue specimen. Recently, we found RCL2®-CS100, a noncross-linking fixative, to display interesting performances regarding tissue morphology and DNA, RNA, and protein quality. We adapted RCL2 tissue fixation protocol so it could be used on a routine and automated laboratory basis, still preserving its good performances. This protocol will be described in detail in the following review.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Delfour, C., Roger, P., Bret, C., Berthe, M. L., Rochaix, P., Kalfa, N., et al. (2006) RCL2, a new fixative, preserves morphology and nucleic acid integrity in paraffin-embedded breast carcinoma and microdissected breast tumor cells. J. Mol. Diagn. 8, 157–169.
Bellet, V., Boissière, F., Bibeau, F., Desmetz, C., Berthe, M. L., Rochaix, P., et al. (2008) Proteomic analysis of RCL2 paraffin-embedded tissues. J. Cell Mol. Med. 12, 2027–2036.
Almeida, A., Paul Thiery J., Magdelénat, H., and Radvanyi, F. (2004) Gene expression analysis by real-time reverse transcription polymerase chain reaction: influence of tissue handling. Anal. Biochem. 328, 101–108.
Bustin, S. A. and Nolan, T. (2004) Pitfalls of quantitative real-time reverse-transcription polymerase chain reaction. J. Biomol. Tech. 15, 155–166.
Copois, V., Bibeau, F., Bascoul-Mollevi, C., Salvetat, N., Chalbos, P., Bareil, C., et al. (2007) Impact of RNA degradation on gene expression profiles: assessment of different methods to reliably determine RNA quality. J. Biotechnol. 127, 549–559.
Fleige, S., Walf, V., Huch, S., Prgomet, C., Sehm, J., and Pfaffl, M. W. (2006) Comparison of relative mRNA quantification models and the impact of RNA integrity in quantitative real-time RT-PCR. Biotechnol. Lett. 28, 1601–1613.
Ho-Pun-Cheung, A., Bascoul-Mollevi, C., Assenat, E., Boissiere-Michot, F., Bibeau, F., Cellier, D., et al. (2009) Reverse transcription-quantitative polymerase chain reaction: description of a RIN-based algorithm for accurate data normalization. BMC Mol. Biol. 15, 10–31.
Strand, C., Enell, J., Hedenfalk, I., and Ferno, M. (2007) RNA quality in frozen breast cancer samples and the influence on gene expression analysis – a comparison of three evaluation methods using microcapillary electrophoresis traces. BMC Mol. Biol. 22, 8–38.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Denouël, A., Boissière-Michot, F., Rochaix, P., Bibeau, F., Boulle, N. (2011). An Alternative Fixative to Formalin Fixation for Molecular Applications: The RCL2®-CS100 Approach. In: Al-Mulla, F. (eds) Formalin-Fixed Paraffin-Embedded Tissues. Methods in Molecular Biology, vol 724. Humana Press. https://doi.org/10.1007/978-1-61779-055-3_19
Download citation
DOI: https://doi.org/10.1007/978-1-61779-055-3_19
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-054-6
Online ISBN: 978-1-61779-055-3
eBook Packages: Springer Protocols