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Image-to-Images Translation for Multiple Virtual Histological Staining of Unlabeled Human Carotid Atherosclerotic Tissue

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Histological analysis of human carotid atherosclerotic plaques is critical in understanding atherosclerosis biology and developing effective plaque prevention and treatment for ischemic stroke. However, the histological staining process is laborious, tedious, variable, and destructive to the highly valuable atheroma tissue obtained from patients.


We proposed a deep learning-based method to simultaneously transfer bright-field microscopic images of unlabeled tissue sections into equivalent multiple sections of the same samples that are virtually stained. Using a pix2pix model, we trained a generative adversarial neural network to achieve image-to-images translation of multiple stains, including hematoxylin and eosin (H&E), picrosirius red (PSR), and Verhoeff van Gieson (EVG) stains.


The quantification of evaluation metrics indicated that the proposed approach achieved the best performance in comparison with other state-of-the-art methods. Further blind evaluation by board-certified pathologists demonstrated that the multiple virtual stains have high consistency with standard histological stains. The proposed approach also indicated that the generated histopathological features of atherosclerotic plaques, such as the necrotic core, neovascularization, cholesterol crystals, collagen, and elastic fibers, are optimally matched with those of standard histological stains.


The proposed approach allows for the virtual staining of unlabeled human carotid plaque tissue images with multiple types of stains. In addition, it identifies the histopathological features of atherosclerotic plaques in the same tissue sample, which could facilitate the development of personalized prevention and other interventional treatments for carotid atherosclerosis.

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The authors would like to acknowledge the instrumental and technical support of multimodal biomedical imaging experimental platform, Institute of Automation, Chinese Academy of Sciences.


This work was supported in part by the National Key Research and Development Program of China under Grant 2017YFA0700401, 2016YFC0103803; the National Natural Science Foundation of China under Grant 81730050, 81827808, 62027901, 81671851, 81527805; CAS Youth Innovation Promotion Association under Grant 2018167; CAS Scientific Instrument R&D Program under Grant YJKYYQ20170075; CAS Key Technology Talent Program; and the Project of High-Level Talents Team Introduction in Zhuhai City (Zhuhai HLHPTP201703).

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Authors and Affiliations



Guanghao Zhang: Conceptualization, Methodology, Software, Writing Original draft.

Bin Ning: Conceptualization, Data acquisition, Writing—original draft.

Hui Hui: Conceptualization, Methodology, Supervision, Writing—review & editing.

Tengfei Yu: Investigation, Data interpretation.

Xin Yang: Conceptualization, Methodology, Data analysis.

Hongxia Zhang: Investigation, Data interpretation.

Jie Tian: Conceptualization, Project administration, Investigation, Supervision.

Wen He: Conceptualization, Project administration, Investigation, Supervision.

Corresponding authors

Correspondence to Jie Tian or Wen He.

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Zhang, G., Ning, B., Hui, H. et al. Image-to-Images Translation for Multiple Virtual Histological Staining of Unlabeled Human Carotid Atherosclerotic Tissue. Mol Imaging Biol 24, 31–41 (2022).

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