GSM9846527: SJNBL033705_D3_3432626_369383; Homo sapiens; OTHER
Fresh Frozen (FF) tissue samples were selected based on stringent RNA quality control metrics, specifically ensuring a DV200 value—representing the percentage of RNA fragments longer than 200 nucleotides—exceeding 30%. Tissue sectioning was performed in accordance with the Visium HD Spatial Gene Expression for FF – Tissue Preparation Guide (Demonstrated Protocol CG000763). Sections of 10 µm thickness were mounted onto Superfrost™ Plus Microscope Slides (Fisherbrand™).The prepared slides were processed using the Visium HD Spatial Gene Expression instrument, which facilitates precise spatial transcriptomic capture. Within the instrument, an additional brightfield image was captured to ensure accurate spatial registration for downstream data analysis. The tissue was then permeabilized, allowing for the transfer of transcriptomic probes onto the Visium HD Spatial Gene Expression capture slide, which features a 6.5 x 6.5 mm capture area. Following probe hybridization and transfer on the Visium HD Spatial Gene Expression slide, subsequent probe extension and sequencing library construction steps were conducted ex situ, adhering to the standard Visium for FFPE workflow (Demonstrated Protocol CG000685). Libraries were indexed utilizing the Dual Index Kit TS, Set A (PN-1000251).Final libraries were sequenced on an Illumina NovaSeq 6000 platform using paired-end dual-indexing with the following read structure: Read 1T—43 cycles; i7 Index—10 cycles; i5 Index—10 cycles; Read 2S—50 cycles. Sequencing was performed at the Hartwell Center, St. Jude Children's Research Hospital, USA.