GSM6449990: Slideseq_T1L_Heart_D7PI; Mus musculus; RNA-Seq
Whole hearts were isolated using aseptic technique and placed in ice cold sterile Hank's Balanced Salt Solution, HBSS.  Blood and other contamination were carefully removed by perfusing the tissues with fresh HBSS. Fresh tissues were immediately embedded in Optimal Cutting Compound (OCT) media and frozen in a liquid-nitrogen-cooled isopentane (EMD Millipore, MX0760) bath for spatial transcriptomics experiments. The tissue blocks were cut into 10µm sections using Thermo Scientific CryoStar NX50 cryostat, and mounted on on a "Curio Seeker Tile (Tile ID #A0004_043, Curio Bioscience). A barcode whitelist and a barcode position file for the corresponding tile were provided by Curio Bioscience. Slide-seq spatial transcriptomics experiment was performed using the Curio Seeker Kit (Curio Bioscience) according to manufacturer's instructions. Briefly, a tissue section from a fresh-frozen reovirus-infected heart at 7 dpi was mounted on a 3mmx3mm spatially indexed bead surface (Curio Seeker Kit, Tile ID #A0004_043, Curio Bioscience). After RNA hybridization and reverse transcription, the tissue section was digested, and the beads were removed from the glass tile and resuspended. Second strand synthesis was then performed by semi-random priming followed by cDNA amplification. A sequencing library was then prepared using the Nextera XT DNA sample preparation kit. The library was sequenced on an Illumina NextSeq 2K (P3 flow cell) using the 100-cycle kit (Read 1 = 50 bp, Read 2 = 80, Index 1 = 10).