GSM6910048: DSP-1001660008975-G-D05; Homo sapiens; OTHER
Briefly, once the slides were deparaffinised and subjected to antigen retrieval procedures, samples were co-incubated with fluorescent-labeled visualization antibodies to detect tumor cells (pan-cytokeratin [CK]), all immune cells (CD45), together with DAPI for nuclei detection. Customizable fluorescent morphology markers, conjugated to unique oligonucleotide tags with an ultraviolet (UV) photocleavable linker, helped visualizing the tissue architecture. The samples were imaged and regions of interest (ROIs) were exposed to UV light that cleaved the linker and released the barcoded oligos for capture by microfluidics and GeoMx DSP then were coupled to next generation sequencing (NGS) readout to profile RNA expression for identifying over 1,800 genes in colon and rectal cancers. The 97 areas of illumination (AOIs) across all slides in mixed stroma/tumor regions was selected After UV illumination, the barcodes were collected in 96-well plates and were used in the NGS readout library preparation procedure protocol. The resulted libraries were sequenced by the Illumina NextSeq 500 platform using 2 x 27 base paired reads