10x Genomics obtained fresh frozen human lymph node from BioIVT Asterand. The tissue was embedded and cryosectioned as described in Visium Spatial Protocols - Tissue Preparation Guide Demonstrated Protocol (CG000240). Tissue sections of 10 µm thickness were placed on Visium Gene Expression Slides.
The H&E image was acquired using a Nikon Ti2-E microscope with the following settings:
- Color camera
- 10X objective
- Numerical Aperture: 0.45
- Exposure: 2 ms
- Gain: 4.5X
The Visium Spatial Gene Expression library was prepared following the Visium Spatial Gene Expression Reagent Kits User Guide (CG000239).
- Sequencing instrument: Illumina NovaSeq 6000
- Sequencing Depth: 123,311 reads per spot
- Sequencing Configuration: 28 x 120 bp
- Sequencing Coverage: Read 1 - 16 bp (includes 16 bp Spatial Barcode, 12 bp UMI); Read 2 - 120 bp (transcript); i7 sample index - 10 bp; i5 sample index - 10 bp.
- Dual-Index set: T1T2-E3
- Slide: V19L01-033
- Area: A1
Key cell metrics were:
- Spots detected under tissue - 4,035
- Median UMI counts per spot - 20,239
- Median genes per spot - 5,999
To maintain donor anonymity, sequence data for this sample is not currently available.