Solvent-free tissue processing using supercritical carbon dioxide
Article first published online: 21 NOV 2012
© 2012 Blackwell Publishing Limited.
Volume 61, Issue 6, pages 1198–1208, December 2012
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
- 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
- supercritical carbon dioxide;
- tissue processing;
- tissue shrinkage;
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.