10x Genomics obtained fresh frozen (FF) mouse brain tissue blocks from Avaden Biosciences. The tissue was sectioned as described in Visium CytAssist Spatial Gene Expression for Fresh Frozen – Tissue Preparation Guide Demonstrated Protocol (CG000636). Tissue section of 10 µm was placed on a standard glass slide, and H&E-stained following methanol fixation. Sections were coverslipped with 85% glycerol, imaged, decoverslipped, followed by destaining as described in Visium CytAssist Spatial Gene Expression for Fresh Frozen – Fixation, H&E Staining, Imaging & Destaining Demonstrated Protocol (CG000614). The glass slide with tissue section was processed via Visium CytAssist instrument to transfer analytes to a Visium CytAssist Spatial Gene Expression slide with 6.5 mm Capture Area following the Visium CytAssist Spatial Gene Expression Reagent Kits for FFPE User Guide (CG000495). RIN Score: 7.0
The H&E image was acquired using Olympus VS200 Slide Scanning Microscope with these settings:
- Olympus Objective magnification: 20x UPLXAPO Objective
- Numerical Aperture: 0.8
- ScopeLED light source: Xcite Novum
- Camera: VS-264C
- Exposure: 500 µs
Libraries were prepared following the Visium CytAssist Spatial Gene Expression Reagent Kits for FFPE User Guide (CG000495).
- Sequencing instrument: Illumina NovaSeq 6, flow cell HLYH7DSX5 (lane 1-4)
- Sequencing depth: 171,410,389 reads / 38.4% saturation
- Sequencing configuration: 28bp read 1 (16bp Visium spatial barcode, 12bp UMI), 90bp read 2 (transcript), 10bp i7 sample barcode and 10bp i5 sample barcode
- Dual-Index set: SI-TS-G12
- Slide: V42U20-064
- Area: A1
Key cell metrics were:
- Spots detected under tissue: 4,298
- Median UMI counts per spot: 19,627
- Median genes per spot: 6,178
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Input Files | Size | md5sum |
---|---|---|
FASTQs | 15.6 GB | a973a66dd59b6c5eb061db81de3b0086 |
Image (TIFF) | 22.2 MB | 9adf9975503d180d4af129fcab831603 |
Manual alignment file | 965 kB | 4c1e882732b1906de66b7600d6ad1c48 |
Probe Set | 2.31 MB | df6ad1d3e39c9406f5a728705d7aad3d |
Microscope image without fiducial frame (TIFF) | 3.36 GB | a7daf74ac02ab71ebc451557e0af36db |
Output Files Format details | Size | md5sum |
---|---|---|
Genome-aligned BAM | 8.6 GB | b26d1eae21cebeb92cb1f8f829c2e5db |
Genome-aligned BAM index | 2.04 MB | 49e451b4a9c5ed6b8bb2934836e8d303 |
Per-molecule read information | 490 MB | 2c49fbee2c7aa3999cba4495e704575e |
Feature / barcode matrix HDF5 (filtered) | 29.8 MB | d343f7d9ef54d4a5454ab6096c2735ef |
Feature / barcode matrix (filtered) | 90.3 MB | bc4a825009ceca52d4b2075fbf67ee30 |
Feature / barcode matrix HDF5 (raw) | 37.5 MB | e36da5a0acaf974c3fcead7bf3b89bfb |
Feature / barcode matrix (raw) | 103 MB | 9a05a29bb5298ef5f1e125107dbaa7e8 |
Clustering analysis | 29.7 MB | f5bff33567e88d2ceeaada2baf7d6848 |
Spatial imaging data | 30.8 MB | 4d3e243f38d620ef5c3d73001a40d8fd |
Summary CSV | 831 B | f975c68ebfe50b90b9cc0334940de10a |
Summary HTML | 5.82 MB | ec5eabf0698562fa84b4c6753a72381c |
Loupe Browser file | 1.81 GB | f558e3d3471e545069818d233ad48050 |