GSM9256396: facial, temporal region, donor 1, section D1; Homo sapiens; OTHER
Fresh frozen human skin tissue was sectioned using a cryostat. Three 5 µm sections were collected for RNA isolation and quality control, and one 5 µm section was mounted on Schott Nexterion™ 3-D Hydrogel-coated slides for spatial transcriptomics. RNA quality was assessed using the Agilent Fragment Analyzer; only samples with RNA quality number (RQN) ≥4 were used. Spatial gene expression libraries were prepared using the 10x Genomics Visium HD Spatial Gene Expression Reagent Kits following the manufacturer's protocols. Tissue sections were formaldehyde-fixed, H&E-stained, scanned, destained, and permeabilized. Whole-transcriptome hybridization was performed overnight using the human probe set v2.0 (18,536 protein-coding genes, 54,018 probes), followed by probe ligation, capture, extension to generate spatially barcoded ligation products, pre-amplification, purification, and library construction. All experimental steps were conducted by Crown Bioscience Germany GmbH.