GSM5388415: Distal lung; Homo sapiens; RNA-Seq
Spatial transcriptomics was performed using Visium Spatial Gene Expression assay (10x Genomics) as per manufacturer's recommendation. Briefly, small pieces of the human lung were incubated in OCT at 4°C for 30 min and frozen in OCT and stored at -80°C.  10 µm sections were placed onto Visium Gateway gene expression slide and stored with desiccants at -80°C until staining and library preparation. Prior to the main experiment, permeabilization time for spatial transcriptomics assay was calculated to be 30 min using the Visium Spatial Tissue Optimization slide by Duke University's Molecular Genomics core facility. H&E staining was performed on the Visium slide and imaged using 10x objective on Keyence BZX800 microscope. Tissue sections were permeabilized immediately to capture RNA molecules onto barcoded spots on the slide. Libraries were then prepared and sequenced (150 bp paired-end) using NovaSeq S4 flow cell. Followed the protocol of 10x Genmics Visium spatial gene expression assay for spatial RNA-seq (3' enrichment)