最近需要处理10x Visium
空间转录组的数据,所以到官网学习了一番基础分析流程的内容。
Slide Parameters
需要搞清楚数据使用的芯片类型,以便后续需要手动对齐图像及跑流程时设置正确的参数:--slide
、--area
、--slidefile
(设备无法联网时需要)。
https://www.10xgenomics.com/support/software/space-ranger/latest/analysis/inputs/image-slide-parameters
Manual Alignment
此步骤为可选项,芯片图片正常的情况下可以直接跳过该步骤,若图片中组织覆盖了四个角的矫正锚点或者遮挡了很多的边框,自动对齐可能不准最好先手动对齐一下图像:
-
Fiducial Alignment
https://www.10xgenomics.com/support/software/space-ranger/latest/tutorials/inputs/image-fiducial-alignment -
CytAssist image alignment
https://www.10xgenomics.com/support/software/space-ranger/latest/analysis/inputs/image-cytassist-image-alignment
Run Pipeline
参考基因组索引构建,人和小鼠可直接下载,需要自己构建可参考:
- Custom References
https://www.10xgenomics.com/support/software/space-ranger/latest/advanced/custom-references
根据实际情况选择合适的流程及参数:
- Spaceranger Count
https://www.10xgenomics.com/support/software/space-ranger/latest/analysis/running-pipelines/space-ranger-count
Review Outputs
所有结果文件:
- https://www.10xgenomics.com/support/software/space-ranger/latest/analysis/outputs/output-overview
网页报告解读:
- https://www.10xgenomics.com/support/software/space-ranger/latest/analysis/outputs/space-ranger-web-summary
空间结果:
- https://www.10xgenomics.com/support/software/space-ranger/latest/analysis/outputs/spatial-outputs
Software Download
https://www.10xgenomics.com/support/software/space-ranger/downloads
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