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Solvent-free tissue processing using supercritical carbon dioxide
Article first published online: 21 NOV 2012
DOI: 10.1111/j.1365-2559.2012.04342.x
© 2012 Blackwell Publishing Limited.
Additional Information
How to Cite
Bleuel, E. P., Roebers, T. P. C., Schulting, E. and den Dunnen, W. F. A. (2012), Solvent-free tissue processing using supercritical carbon dioxide. Histopathology, 61: 1198–1208. doi: 10.1111/j.1365-2559.2012.04342.x
Publication History
- Issue published online: 28 NOV 2012
- Article first published online: 21 NOV 2012
- Accepted manuscript online: 6 JUN 2012 12:47PM EST
- Date of submission 13 February 2012 Accepted for publication 4 June 2012
Keywords:
- dehydration;
- paraffination;
- solvent-free;
- supercritical carbon dioxide;
- tissue processing;
- tissue shrinkage;
- xylene-free
Bleuel E P, Roebers T P C, Schulting E & den Dunnen W F A (2012) Histopathology Solvent-free tissue processing using supercritical carbon dioxide
Aims: Xylene is most often employed in tissue processing protocols for paraffin embedding, but poses a health hazard. The aim of this study was to evaluate a solvent-free processing protocol that uses supercritical carbon dioxide (scCO2) as an intermediate.
Methods and Results: A series of tests (with bovine tissues) was run, evaluating dehydration and tissue shrinkage in our new scCO2-based protocol as compared with routine processing using a graded ethanol and xylene series. A series of tests was then run to evaluate the significance of processing parameters for the outcome. Finally, a validation series was performed with optimal conditions, testing various human tissues with several staining methods. The tissue water content after paraffination was the same with our new scCO2-based protocol and the routine xylene-based protocol. Tissue shrinkage was similar with the two methods, at ∼15%, which is also similar to values in the literature. In the validation series, the human tissues showed good morphology with strong staining, probably because of stronger antigenicity.
Conclusions: This scCO2-based protocol has been shown to be a good solvent-free, alternative form of tissue processing. Although not the focus of this article, the time needed for tissue processing with this new protocol is within 4 h, and there is no need to change macroscopy/sectioning protocols.