GSM7993713: ACC11_spatial; Homo sapiens; OTHER
After cutting normal adrenal tissue of Normal1_spatial, Normal2_spatia and Normal3_spatia and adrenocortical carcinoma of ACC11_spatial, ACC12_spatial, ACC13_spatial and ACC14_spatial into 4-5 mm3 with a dust-free scalpel, dust-free paper was taken to wipe the blood stains from the tissue surface and then fresh tissue was embedded with OCT and placed in -80°C for rapid freezing. The tissues were frozen sectioned (10 µm) on a Lycra frozen sectioning machine and the sections were attached to ST microarrays. Then, HE staining were performed by dehydration with isopropyl alcohol for 1 min. And bright-field images were taken on a 3D HISTECH Pannoramic MIDI FL full-slide scanner at 20X resolution. Tissue sections were placed in the corresponding capture zones on 10X Visium tissue permeabilization optimization slides. The slides for optimization experiments have 8 capture regions, and by removing the positive and negative controls from them, 6 additional sections are needed for permeabilization optimization experiments. The sections used for optimization are attached to different capture regions and then treated with permeabilizing enzymes for different times. The mRNA released during the permeabilization process binds to primers with poly-dT on the slides to generate fluorescently labeled cDNA. finally, the excess tissue is removed, leaving the fluorescently labeled cDNA to be observed using a fluorescence microscope to determine the optimal permeabilization time. Tissue sections placed in the capture region on gene expression slides are fixed and stained, permeabilized, and cellular mRNA is captured by primers at the gene expression site. The primers capture mRNA at specific sites to generate all cDNAs with a common spatial barcode. Libraries are generated from the cDNAs and sequenced, and the spatial barcodes are then used to correlate read information with tissue section images for spatial gene expression mapping. 10X Visium