Step 1: Prepare Your Sample
Embed, section, and place fresh-frozen tissue onto a capture area of the gene expression slide. Each capture area has thousands of barcoded spots, each containing millions of capture oligonucleotides with spatial barcodes unique to that spot.
Step 2: Stain and Image the Tissue
Step 3: Permeabilise Tissue and Construct Library
Permeabilise the tissue to release mRNA from the cells, which binds to the spatially-barcoded oligonucleotides present on the spots. A reverse transcription reaction produces cDNA from the captured mRNA. The barcoded cDNA is then pooled for downstream processing to complete a sequencing-ready library.
Step 4: Sequence
The resulting 10x Barcoded library is compatible with standard NGS short-read sequencing on Illumina sequencers for massive transcriptional profiling of entire tissue sections.
Step 5: Analyse and Visualise Your Data
Use the Space Ranger analysis software to process your Spatial Gene Expression data and interactively explore the results with the Loupe Browser visualisation software.