GSM8659692: Adrenal gland; Mus musculus; OTHER
Regions of interest (ROIs) within the mouse adrenal glands were identified based on their morphology, orientation (determined through H&E staining), and RNA quality assessment via DV200, measured using an Agilent 2100 Bioanalyzer. The FFPE tissue blocks were then sectioned into 5 μm slices using a Leica RM2255 microtome. Total RNA was extracted from the FFPE tissue using the RNeasy FFPE kit (73504, Qiagen).  FFPE Visium Spatial Gene Expression analysis (10x Genomics) was carried out on a CytAssist instrument following the manufacturer's instructions. Briefly, tissue sections were mounted on SuperFrost™ Plus Microscope Slides (#22-037-246, FisherScientific) and left to dry overnight. The slides were then incubated at 60°C for 2 hours before undergoing deparaffinization. or the H&E-stained sections, decrosslinking was performed immediately, followed by the incorporation of probe panels targeting the entire mouse transcriptome. After the probe pairs hybridized with their corresponding target transcripts, the slides were processed on the Visium CytAssist instrument for permeabilization and RNase treatment. Subsequently, transcript capture was conducted, and the Visium Library Preparation protocol from 10x Genomics was followed. Four cDNA libraries were diluted and pooled to a final concentration of 650 pM in a 20 μl volume, then sequenced using a P1 flow cell on the Illumina NextSeq 2000 platform.
GEO
GSM8659692_r1