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Solvent‐free tissue processing using supercritical carbon dioxide
Author(s) -
Bleuel Erik P,
Roebers Thijs P C,
Schulting Edwin,
den Dunnen Wilfred F A
Publication year - 2012
Publication title -
histopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.626
H-Index - 124
eISSN - 1365-2559
pISSN - 0309-0167
DOI - 10.1111/j.1365-2559.2012.04342.x
Subject(s) - xylene , solvent , supercritical carbon dioxide , supercritical fluid , biomedical engineering , chemistry , chromatography , medicine , organic chemistry , benzene
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 (scCO 2 ) as an intermediate. Methods and Results: A series of tests (with bovine tissues) was run, evaluating dehydration and tissue shrinkage in our new scCO 2 ‐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 scCO 2 ‐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 scCO 2 ‐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.