Spatial polyA+ RNAseq: Sample VM3-1_Lumbar
Human DRGs were collected within 4 hrs of cross-clamp from neurologically dead organ donors, and immediately fresh frozen in dry ice. DRGs were cryosectioned onto SuperFrost Plus charged slides at 10µm. To sample a larger subset of neurons, two sections were utilized from each donor and each 10µm section was 200µm apart. Visium tissue optimization and spatial gene expression protocols were followed exactly as described by 10x Genomics (https://www.10xgenomics.com/) using Haematoxylin and Eosin as the counterstain. Optimal permeabilization time was obtained at 12 min incubation with permeabilization enzyme. mRNA library preparation and sequencing (Illumina Nextseq 500) were done at the Genome Center in the University of Texas at Dallas Research Core Facilities.