Conventional xylene and xylene-free methods for routine histopathological preparation of tissue sections.
Author(s): Metgud R, Astekar MS, Soni A, Naik S, Vanishree M
Publication: Biotech Histochem, 2013, Vol. 88, Page 235-41
PubMed ID: 23373510 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to determine the effects of clearing agent used during processing and staining of sections on morphological preservation in formalin-fixed paraffin-embedded (FFPE) tissue.
Conclusion of Paper
Xylene versus xylene-free tissue processing and H&E staining methods did not significantly affect the morphological preservation of FFPE odontogenic cysts and squamous cell carcinomas.
Studies
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Study Purpose
The purpose of this study was to determine the effects of clearing agent used during processing and staining of FFPE sections on morphological preservation. A total of 30 specimens were included in the study, half of which were processed using a conventional xylene method (formalin, dehydration in graded alcohol, clearing with xylene, and embedding in wax) while the other half were processing using a xylene-free method (formalin, water, isopropanol, and embedding in paraffin). The specimens processed using the xylene-free method also underwent xylene-free H&E staining (deparaffinization, clearing in soap water, rehydration in graded alcohol, H&E staining) while the conventionally processed specimens used xylene for clearing during H&E staining of sections.
Summary of Findings:
There were no significant differences in specimen shrinkage, quality of tissue or staining, tissue architecture, or integrity of epithelial, glandular, fibrous, muscle, adipose, vascular, or nerve tissue, or inflammatory cells between the xylene and xylene-free processing and staining methods as evaluated blindly by three pathologists on a 0-3 scale. However, tissue shrinkage was insignificantly greater and the quality of morphological preservation was insignificantly better when tissue was processed and stained using xylene compared to xylene-free methods.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Neoplastic - Carcinoma
- Neoplastic - Benign
Platform:
Analyte Technology Platform Morphology H-and-E microscopy Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Clearing agent Xylene
Liquid dishwashing soap in water
Isopropanol