GSM7806338: C_DAA003_7; Homo sapiens; OTHER
Normal adrenal glands were dissected and placed into histidine tryptophan ketoglutarate (HTK) solution. Within 2 hours of receiving the tissue, the gland was cut into sections for fresh frozen tissue and to create OCT blocks for storage. 10-micron sections of OCT blocks were cut and used for H&E to ensure a well-preserved tissue to use in further experiments. Adjacent sections were cut and used for spatial transcriptomics and immunohistochemical analysis. Frozen samples were OCT embedded and sectioned at 10μm on a Cryostar NX70 cryostat (Thermo Fisher Scientific). Capture sections were fixed with methanol, stained with H&E, and imaged on an Evos M7000 (ThermoFisher) with brightfield settings. Capture sections were permeabilized and processed to generate RNA libraries following the 10x Visium protocol. Libraries were sequenced to a depth of 60,000 read pairs per spot calculated from the image, on a Novaseq6000 (Illumina) sequencer with 151x151 bp runs.