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Stain–Decolorize–Stain (SDS): a new technique for multiple staining

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Abstract

Multiple staining of more than one gene/antigen on a single tissue section is an indispensable tool in cell and tissue research. However, most of the available multiple staining techniques have limitations, and there has been no technique to simultaneously visualize and distinguish tissue antigens, nucleotide sequences and other chemical compounds on the same slide. Here, we present a practical and economic multiple stain technique, with which multiple cellular components including mRNA (with in situ hybridization), antigen epitope (with immunohistochemistry) and chemical molecules (with histochemistry) can be stained on a single tissue section to study their relationship. In addition, this technique also offers the possibility to evaluate morphology with an H&E staining on the same sections. We used the placenta, pancreas, breast ductal carcinoma, colon adenocarcinoma, cerebellum, tonsil and heart tissue sections to evaluate the applicability of this new technique. The sensitivity and specificity of the technique have been tested, and an optimal protocol is recommended. Its applications in surgical pathology and research are discussed. This technique offers a novel tool to evaluate the relationship among multiple components at the same or adjacent locations to meet the needs of pathology diagnosis and research.

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Acknowledgments

We thank Lu Yao, Na Niu, Zhuo Li, Yuxuan Liu, Yong Guo and Qi Cao from Department of Pathology of Peking University Health Science Center and Yiqun Geng, Tao Huang and Li Du from Department of Pathology of Shantou University Medical College for providing the tissue samples, comments and suggestions.

Conflict of interest

The authors declared no conflict of interest with respect to the research, authorship, and publication of this article. This work was supported by grants from the National Nature Science Foundation of China (No. 81030033, 30971150 to J.G.).

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Correspondence to Jiang Gu.

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Jing Li and Yan Zhou contributed equally to this work.

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Li, J., Zhou, Y. & Gu, J. Stain–Decolorize–Stain (SDS): a new technique for multiple staining. Histochem Cell Biol 141, 251–262 (2014). https://doi.org/10.1007/s00418-013-1177-7

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  • DOI: https://doi.org/10.1007/s00418-013-1177-7

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