GSM8192638: DRG, round1, s5; Sus scrofa; OTHER
Methanol fixation and Eosin and Haematoxylin staining for an initial tissue quality control analysis was performed as described in the 10XGenomics Methanol Fixation, H&E Staining & Imaging for Visium Spatial Protocols (Demonstrated Protocol CG000160). We stained a total of 60 sections, each 10 µm in thickness, from 22 fresh frozen DRG obtained from 7 female pigs. Different permeabilization times (3, 6, 12, 18, 24 and 30 minutes) were evaluated following the 10XGenomics Visium Spatial Gene Expression Reagent Kits - Tissue Optimization User Guide CG000238 Rev E. This protocol required the reagents from the Visium Spatial Tissue Optimization Reagent Kit, PN-1000192 (stored at −20°C) and the Visium Spatial Tissue Optimization Slide Kit PN-1000191 (stored at ambient temperature). The exposure time of the permeabilization enzyme was 18 minutes based on the analysis of the tissue optimization experiment. We used the spatial sequencing protocol Visium Spatial Gene Expression Reagent Kits, 16 rxns PN-1000186 and Library Construction Kits, 16 rxns PN-1000190. All reagents were stored at -20°C. Additionally, Visium Spatial Gene Expression Slide Kits, 16 rxns PN-1000185 were acquired and stored at ambient temperature. The protocol was performed exactly as stated in the Visium Spatial Gene Expression Reagents Kits User Guide CG000239 Rev F.