Research Papers: General

Investigation of influences of the paraformaldehyde fixation and paraffin embedding removal process on refractive indices and scattering properties of epithelial cells

[+] Author Affiliations
Jing-Wei Su

National Taiwan University, Institute of Biomedical Electronics and Bioinformatics, Taipei 10617, Taiwan

Wei-Chen Hsu

National Taiwan University, Institute of Biomedical Electronics and Bioinformatics, Taipei 10617, Taiwan

Jeng-Wei Tjiu

National Taiwan University Hospital, Department of Dermatology, Taipei 10617, Taiwan

Chun-Pin Chiang

National Taiwan University Hospital, Department of Dentistry, Taipei 10617, Taiwan

Chao-Wei Huang

National Taiwan University, Department of Animal Science and Technology, Taipei 10617, Taiwan

Kung-Bin Sung

National Taiwan University, Institute of Biomedical Electronics and Bioinformatics, Taipei 10617, Taiwan

National Taiwan University, Department of Electrical Engineering, Taipei 10617, Taiwan

National Taiwan University, Molecular Imaging Center, Taipei 10617,Taiwan

J. Biomed. Opt. 19(7), 075007 (Jul 28, 2014). doi:10.1117/1.JBO.19.7.075007
History: Received April 8, 2014; Revised June 18, 2014; Accepted June 30, 2014
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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 450cm1, respectively, estimated from paraffin-embedded oral epithelial tissue after restoration of RI and volume.

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© 2014 Society of Photo-Optical Instrumentation Engineers

Citation

Jing-Wei Su ; Wei-Chen Hsu ; Jeng-Wei Tjiu ; Chun-Pin Chiang ; Chao-Wei Huang, et al.
"Investigation of influences of the paraformaldehyde fixation and paraffin embedding removal process on refractive indices and scattering properties of epithelial cells", J. Biomed. Opt. 19(7), 075007 (Jul 28, 2014). ; http://dx.doi.org/10.1117/1.JBO.19.7.075007


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