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Journal of Biomedical Optics

Investigation of influences of the paraformaldehyde fixation and paraffin embedding removal process on refractive indices and scattering properties of epithelial cells
Author(s): Jing-Wei Su; Wei-Chen Hsu; Jeng-Wei Tjiu; Chun-Pin Chiang; Chao-Wei Huang; Kung-Bin Sung
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Paper Abstract

The scattering properties and refractive indices (RI) of tissue are important parameters in tissue optics. These parameters can be determined from quantitative phase images of thin slices of tissue blocks. However, the changes in RI and structure of cells due to fixation and paraffin embedding might result in inaccuracies in the estimation of the scattering properties of tissue. In this study, three-dimensional RI distributions of cells were measured using digital holographic microtomography to obtain total scattering cross sections (TSCS) of the cells based on the first-order Born approximation. We investigated the slight loss of dry mass and drastic shrinkage of cells due to paraformaldehyde fixation and paraffin embedding removal processes. We propose a method to compensate for the correlated changes in volume and RI of cells. The results demonstrate that the TSCS of live cells can be estimated using restored cells. The percentage deviation of the TSCS between restored cells and live cells was only −8%. Spatially resolved RI and scattering coefficients of unprocessed oral epithelium ranged from 1.35 to 1.39 and from 100 to 450  cm−1, respectively, estimated from paraffin-embedded oral epithelial tissue after restoration of RI and volume.

Paper Details

Date Published: 28 July 2014
PDF: 9 pages
J. Biomed. Opt. 19(7) 075007 doi: 10.1117/1.JBO.19.7.075007
Published in: Journal of Biomedical Optics Volume 19, Issue 7
Show Author Affiliations
Jing-Wei Su, National Taiwan Univ. (Taiwan)
Wei-Chen Hsu, National Taiwan Univ. (Taiwan)
Jeng-Wei Tjiu, National Taiwan Univ. Hospital (Taiwan)
Chun-Pin Chiang, National Taiwan Univ. Hospital (Taiwan)
Chao-Wei Huang, National Taiwan Univ. (Taiwan)
Kung-Bin Sung, National Taiwan Univ. (Taiwan)

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