title: GSM7669695: WT.DDC.007.Periportal.IBA1Posi; Mus musculus; OTHER
library_name: GSM7669695
library_construction_protocol: Samples were incubated with DNA-oligo barcoded RNA-ISH probes which were conjugated with a UV-photocleavable linker following standard ISH protocols, along with fluorescence labeled antibodies for visualization of morphological structures. Regions of interest (ROI) are selected on the visualized tissue. Using fluorescent morphology markers and auto-segmentation algorithm, individual segments was divided. The photocleaved oligos were aspirated via a microcapillary tube and stored in microplate wells by the GeoMx® Digital Spatial Profiler (DSP) platform. For more information about DSP protocols please see Merritt et al. Nature Biotech 2020 (https://doi.org/10.1038/s41587-020-0472-9) Each collection of oligo tags from one well (representing an AOI from the tissue section) was indexed with i7xi5 unique dual indexes using GeoMx SeqCode primers with 18 cycles of PCR. After PCR, indexed AOIs were pooled and purified in two rounds of AMPure XP PCR purification using 1.2x bead:sample ratio.