GSM8819276: NCCLC5; Homo sapiens; OTHER
We received fresh-frozen tissue samples that had been stored in the National Cancer Center biobank, embedded in OCT, and stored at -80°C. Tissue block was cut into 10-μm sections, which were analyzed with a Visium Spatial Gene Expression Kit 3' v1 (10x Genomics) according to the manufacturer's instructions in the Visium Spatial Gene Expression Reagent Kit User Guide(﻿10X Genomics). The optimal permeabilization time was 24 min. Sliced tissue pieces were stained with H&E, imaged using a BZ-X710 microscope (Keyence, Osaka, Japan) under a 20 x objective, and processed for spatial transcriptomics. After permeablization, mRNA released from the overlying cells is captured by the primers on the spots. Then spatially barcoded cDNA is synthesied from mRNA. Library preparation was performed included cDNA amplification, fragmentation, hearing and 5′ adaptor and sample index attachment. Sequencing was performed on a DNBSEQ (MGI Tech, Shenzhen, China).