10x Visium空转数据基础流程还得看官网资料,看完事半功倍

随笔2周前发布 安妮
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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

10x Visium空转数据基础流程还得看官网资料,看完事半功倍

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:

10x Visium空转数据基础流程还得看官网资料,看完事半功倍

  1. Fiducial Alignment
    https://www.10xgenomics.com/support/software/space-ranger/latest/tutorials/inputs/image-fiducial-alignment

  2. 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:

  1. 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:

  1. 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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