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miRNA in situ hybridization

XD Xinyue Du
SW Shuangmiao Wang
XL Xingyan Liu
TH Tao He
XL Xiangui Lin
SW Simin Wu
DW Dan Wang
JL Jiao Li
WH Wenhua Huang
HY Huiling Yang
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Lung adenocarcinoma tissue chip was dewaxed, rehydrated through a descending ethanol gradient, then treated with 0.1% hydrochloric acid–ethanol for 15 min. The tissue chip was incubated with diluted pepsin in 3% fresh citrate buffer at 37 °C for 20 min and then washed with phosphate-buffered saline (PBS). The chip was fully digested to expose the mRNA of the tissue on the chip, which can enhance the subsequent immune hybridization signal. The digoxin-labeled miRNA probe (Biosense Bioscience Co. Ltd., Guangzhou, China) was diluted with the hybridization diluent and hybridized overnight at 37 °C. Blocking solution was added dropwise, DAB staining solution was used for color development, and hematoxylin counterstaining was performed. Slides were dehydrated, dried at room temperature, and sealed with neutral resin.

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