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How thick are the paraffin-embedded tissue sections routinely prepared in laboratory? A morphometric study using a
T Masuda1, J Kawaguchi, H Oikawa
1Department of Pathology, School of Medicine, Iwate Medical University, Morioka, Japan. masuda@iwate-med.ac.jp
Pathology International
|May 20, 1998
Summary
The actual thickness of formalin-fixed, paraffin-embedded tissue sections often differs from the cut thickness, impacting measurements. These variations introduce significant errors in quantitative analyses of tissue sections.
Area of Science:
- Histology
- Microscopy
- Biomedical Engineering
Background:
- Accurate tissue section thickness is crucial for quantitative analysis in histology.
- Formalin-fixed, paraffin-embedded (FFPE) tissues are widely used in research and diagnostics.
- Variations in section thickness can lead to errors in stereological measurements.
Purpose of the Study:
- To evaluate the discrepancies between the intended cut thickness and the actual measured thickness of FFPE tissue sections.
- To quantify the impact of these thickness variations on potential quantitative analyses.
Main Methods:
- FFPE liver tissue blocks from Wistar rats were sectioned at predetermined thicknesses (3-18 microns).
- Confocal laser scanning microscopy was employed for observation.
- Section thickness was measured using specialized microscope software analyzing vertically cut images.
Main Results:
- Measured section thicknesses consistently deviated from the preselected cut thicknesses.
- Mean measured thicknesses ranged from 4.3 microns (for 3-micron cuts) to 17.7 microns (for 18-micron cuts).
- Surface irregularities and inclination of sections contributed to thickness variations, leading to standard errors up to 10% in evaluated parameters.
Conclusions:
- The difference between cut and measured thicknesses of FFPE sections is significant.
- These variations introduce considerable error into quantitative analyses like volume and surface densities.
- Researchers must account for these inherent thickness variations in FFPE sections for accurate stereological evaluations.