Influence of hematoxylin and eosin staining on linear birefringence measurement of fibrous tissue structures in polarization microscopy

Liangyu Deng; Chunyan Chen; Wenxi Yu; Conghui Shao; Zan Shen; Yonggang Wang; Chao He; Hongtao Li; Zhiyan Liu; Honghui He; Hui Ma

doi:10.1117/1.JBO.28.10.102909

30 September 2023 Influence of hematoxylin and eosin staining on linear birefringence measurement of fibrous tissue structures in polarization microscopy

Liangyu Deng, Chunyan Chen, Wenxi Yu, Conghui Shao, Zan Shen, Yonggang Wang, Chao He, Hongtao Li, Zhiyan Liu, Honghui He, Hui Ma

Author Affiliations +

Journal of Biomedical Optics, Vol. 28, Issue 10, 102909 (September 2023). https://doi.org/10.1117/1.JBO.28.10.102909

Abstract

Significance

For microscopic polarization imaging of tissue slices, two types of samples are often prepared: one unstained tissue section for polarization imaging to avoid possible influence from staining dyes quantitatively and one hematoxylin-eosin (H&E) stained adjacent tissue section for histological diagnosis and structural feature identification. However, this sample preparation strategy requires high-quality adjacent tissue sections, and labeling the structural features on unstained tissue sections is impossible. With the fast development of data driven-based polarimetric analysis, which requires a large amount of pixel labeled images, a possible method is to directly use H&E stained slices, which are standard samples archived in clinical hospitals for polarization measurement.

Aim

We aim to study the influence of hematoxylin and eosin staining on the linear birefringence measurement of fibrous tissue structures.

Approach

We examine the linear birefringence properties of four pieces of adjacent bone tissue slices with abundant collagen fibers that are unstained, H&E stained, hematoxylin (H) stained, and eosin (E) stained. After obtaining the spatial maps of linear retardance values for the four tissue samples, we carry out a comparative study using a frequency distribution histogram and similarity analysis based on the Bhattacharyya coefficient to investigate how H&E staining affects the linear birefringence measurement of bone tissues.

Results

Linear retardance increased after H&E, H, or E staining (41.7%, 40.8%, and 72.5% increase, respectively). However, there is no significant change in the imaging contrast of linear retardance in bone tissues.

Conclusions

The linear retardance values induced by birefringent collagen fibers can be enhanced after H&E, H, or E staining. However, the structural imaging contrasts based on linear retardance did not change significantly or the staining did not generate linear birefringence on the sample area without collagen. Therefore, it can be acceptable to prepare H&E stained slices for clinical applications of polarimetry based on such a mapping relationship.

CC BY: © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 International License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

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Liangyu Deng, Chunyan Chen, Wenxi Yu, Conghui Shao, Zan Shen, Yonggang Wang, Chao He, Hongtao Li, Zhiyan Liu, Honghui He, and Hui Ma "Influence of hematoxylin and eosin staining on linear birefringence measurement of fibrous tissue structures in polarization microscopy," Journal of Biomedical Optics 28(10), 102909 (30 September 2023). https://doi.org/10.1117/1.JBO.28.10.102909

Received: 26 April 2023; Accepted: 19 September 2023; Published: 30 September 2023

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