Application of RNAscope Multi-Channel Fluorescence Detection in Organoid Studies Based on Alveolar Epithelial Type II Cells

SUN Jianhui¹^,2, LIU Di¹^,2, ZHANG Huacai¹^,2, ZHANG Anqiang¹^,2, MI Junwei¹^,2, JIANG Jianxin¹^,2*, ZENG Ling¹^,2*

(¹Department of Trauma Medical Center, Daping Hospital, Army Medical University, Chongqing 400042, China; ²State Key Laboratory of Trauma, Burns and Combined Injuries, Chongqing 400042, China)

Abstract:

This study aims to investigate the application of RNAscope multi-channel fluorescence detection in organoids research and establish relative experimental system. AEC2s (alveolar epithelial type II cells) were sorted by magnetic beads from SD rat and identified their purity by immunofluorescence. The isolated AEC2s cells were cultured in lung organoids. At 15 days, inverted microscope was detected to study the growth law at 15 days, and HE staining was also performed to show the morphological structure of organoids. RNAscope multi-channel fluorescence detection was performed to determine the expression of Sftpc, Aqp5, krt5 and ki67 in single cells of lung organoids in situ. The results showed that RNAscope multichannel fluorescence detection could be used to analyse the expression of multiple RNA at the single-cell level in organoids simultaneously, and to provide complete histomorphological information while obtaining RNAdatabase in single cells, which improved the understanding of the complex biological correlation between diseases and markers. Therefore, RNAscope multi-channel fluorescence detection propose a new method for the study of organoids at the single-cell level.

CSTR: 32200.14.cjcb.2021.10.0005