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- <h1 id="seo-header">【复习】单细胞最佳实践的流程</h1>
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- <h2 id="配置环境">配置环境</h2>
- <ul>
- <li><a href="/-ji-lu--an-zhuang-sheng-xin-de-dai-ma-bian-xie-huan-jing">基础编程环境</a></li>
- <li><a href="/-fu-ke-GitHub-wen-jian-jia-su">GitHub 下载加速</a></li>
- </ul>
- <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br></pre></td><td class="code"><pre><code class="hljs R">options<span class="hljs-punctuation">(</span>BioC_mirror<span class="hljs-operator">=</span><span class="hljs-string">"https://mirrors.ustc.edu.cn/bioc/"</span><span class="hljs-punctuation">)</span> <span class="hljs-comment">## 指定 BiocManager::install 镜像</span><br>options<span class="hljs-punctuation">(</span><span class="hljs-string">"repos"</span> <span class="hljs-operator">=</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span>CRAN<span class="hljs-operator">=</span><span class="hljs-string">"https://mirrors.tuna.tsinghua.edu.cn/CRAN/"</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span> <span class="hljs-comment">## 指定 install.packages 镜像,</span><br>options<span class="hljs-punctuation">(</span>ggrepel.max.overlaps <span class="hljs-operator">=</span> <span class="hljs-literal">Inf</span><span class="hljs-punctuation">)</span><br></code></pre></td></tr></table></figure>
- <figure class="highlight bash"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br><span class="line">5</span><br><span class="line">6</span><br><span class="line">7</span><br><span class="line">8</span><br><span class="line">9</span><br><span class="line">10</span><br><span class="line">11</span><br><span class="line">12</span><br><span class="line">13</span><br><span class="line">14</span><br><span class="line">15</span><br><span class="line">16</span><br><span class="line">17</span><br><span class="line">18</span><br><span class="line">19</span><br><span class="line">20</span><br><span class="line">21</span><br><span class="line">22</span><br><span class="line">23</span><br><span class="line">24</span><br><span class="line">25</span><br><span class="line">26</span><br><span class="line">27</span><br><span class="line">28</span><br><span class="line">29</span><br><span class="line">30</span><br><span class="line">31</span><br><span class="line">32</span><br><span class="line">33</span><br><span class="line">34</span><br><span class="line">35</span><br><span class="line">36</span><br><span class="line">37</span><br><span class="line">38</span><br><span class="line">39</span><br><span class="line">40</span><br><span class="line">41</span><br><span class="line">42</span><br><span class="line">43</span><br></pre></td><td class="code"><pre><code class="hljs bash">nano ~/.Rprofile <span class="hljs-comment"># 内容如上,配置镜像</span><br>conda create -n seurat5 -c conda-forge r-seurat=4.3.0 -y<br>conda activate seurat5<br>conda install -c conda-forge r-irkernel -y<br>conda install -c conda-forge r-devtools -y<br>conda install -c conda-forge r-biocmanager -y<br>conda install -c conda-forge r-tidyverse -y<br>conda install -c conda-forge r-clustree -y<br>Rscript -e <span class="hljs-string">"IRkernel::installspec(name='seurat5', displayname='seurat5')"</span><br><br><span class="hljs-comment">## 开始 seurat5 的安装</span><br>wget https://github.com/satijalab/seurat/archive/refs/heads/seurat5.zip -O seurat5.zip<br>Rscript -e <span class="hljs-string">"devtools::install_local('seurat5.zip')"</span><br><span class="hljs-comment">## Rscript -e "devtools::install_local('seurat5.zip', host = 'xxx.limour.top/token/api.github.com')"</span><br><span class="hljs-comment">## options(Seurat.object.assay.version = 'v5') 使用 seurat5</span><br>conda install -c conda-forge hdf5 -y<br>wget https://github.com/bnprks/BPCells/archive/refs/heads/main.zip -O BPCells.zip<br>Rscript -e <span class="hljs-string">"devtools::install_local('BPCells.zip')"</span><br><br><span class="hljs-comment">## 开始 DoubletFinder 的安装</span><br>wget https://github.com/chris-mcginnis-ucsf/DoubletFinder/archive/refs/heads/master.zip -O DoubletFinder.zip<br>Rscript -e <span class="hljs-string">"devtools::install_local('DoubletFinder.zip')"</span><br><br><span class="hljs-comment">## 开始 decontX 的安装</span><br>wget https://github.com/campbio/decontX/archive/refs/heads/main.zip -O decontX.zip<br>conda install -c conda-forge imagemagick -y<br>conda install -c conda-forge r-ggrastr -y<br>Rscript -e <span class="hljs-string">'BiocManager::install("celda")'</span><br>Rscript -e <span class="hljs-string">"devtools::install_local('decontX.zip')"</span><br><span class="hljs-comment">## api.github.com 错误就换用下面的命令</span><br><span class="hljs-comment"># Rscript -e 'remotes::install_local("decontX.zip", repos = NULL, type = "source")'</span><br><span class="hljs-comment"># Rscript -e "devtools::install_local('decontX.zip', repos = NULL, type='source')"</span><br><span class="hljs-comment"># Rscript -e 'install.packages("decontX.zip", repos = NULL, type = "source")'</span><br><br><span class="hljs-comment">## 开始 plot1cell 的安装</span><br>wget https://github.com/mojaveazure/loomR/archive/refs/heads/master.zip -O loomR.zip<br>Rscript -e <span class="hljs-string">"devtools::install_local('loomR.zip')"</span><br>wget https://github.com/HaojiaWu/plot1cell/archive/refs/heads/master.zip -O plot1cell.zip<br>Rscript -e <span class="hljs-string">'BiocManager::install(c("biomaRt","EnsDb.Hsapiens.v86","GEOquery","simplifyEnrichment"))'</span><br>Rscript -e <span class="hljs-string">"devtools::install_local('plot1cell.zip')"</span><br><br><span class="hljs-comment">## 开始 SCTransform 的安装</span><br>Rscript -e <span class="hljs-string">'BiocManager::install("glmGamPoi")'</span><br></code></pre></td></tr></table></figure>
- <h3 id="附加-安装SCP">附加 安装SCP</h3>
- <figure class="highlight bash"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br><span class="line">5</span><br><span class="line">6</span><br><span class="line">7</span><br><span class="line">8</span><br><span class="line">9</span><br><span class="line">10</span><br><span class="line">11</span><br><span class="line">12</span><br><span class="line">13</span><br><span class="line">14</span><br><span class="line">15</span><br><span class="line">16</span><br></pre></td><td class="code"><pre><code class="hljs bash">conda create -n r-scp -c conda-forge r -y<br>conda activate r-scp<br>conda install -c conda-forge r-irkernel -y<br>conda install -c conda-forge r-devtools -y<br>conda install -c conda-forge r-biocmanager -y<br>Rscript -e <span class="hljs-string">"IRkernel::installspec(name='r-scp', displayname='r-scp')"</span><br><span class="hljs-comment">## 开始 SCP 的安装</span><br>wget https://github.com/zhanghao-njmu/SCP/archive/refs/heads/main.zip -O SCP.zip<br>conda install -c conda-forge r-ggplot2 -y<br>conda install -c conda-forge r-gridgraphics -y<br>Rscript -e <span class="hljs-string">'BiocManager::install("ggtree")'</span><br>Rscript -e <span class="hljs-string">"devtools::install_local('SCP.zip')"</span><br><br><span class="hljs-comment">## 找到 conda 的路径</span><br>whereis conda<br><span class="hljs-comment"># /opt/conda/bin/conda</span><br></code></pre></td></tr></table></figure>
- <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br><span class="line">5</span><br><span class="line">6</span><br></pre></td><td class="code"><pre><code class="hljs R"><span class="hljs-comment">## 创建 SCP 的 python 环境</span><br>options<span class="hljs-punctuation">(</span>reticulate.conda_binary <span class="hljs-operator">=</span> <span class="hljs-string">"/opt/conda/bin/conda"</span><span class="hljs-punctuation">,</span> SCP_env_name <span class="hljs-operator">=</span> <span class="hljs-string">"python-scp"</span><span class="hljs-punctuation">)</span><br>SCP<span class="hljs-operator">::</span>PrepareEnv<span class="hljs-punctuation">(</span><br> miniconda_repo <span class="hljs-operator">=</span> <span class="hljs-string">"https://mirrors.bfsu.edu.cn/anaconda/miniconda"</span><span class="hljs-punctuation">,</span><br> pip_options <span class="hljs-operator">=</span> <span class="hljs-string">"-i https://pypi.tuna.tsinghua.edu.cn/simple"</span><br><span class="hljs-punctuation">)</span><br></code></pre></td></tr></table></figure>
- <h2 id="准备数据">准备数据</h2>
- <ul>
- <li><a href="/er-dai-ce-xu-shu-ju-chu-li-zhi-shu-ju-ge-shi-shuo-ming">二代测序数据格式</a></li>
- <li><a href="/cellranger-ding-liang--One-Library--Multiple-Flowcells">cellranger 定量</a></li>
- </ul>
- <figure class="highlight bash"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br></pre></td><td class="code"><pre><code class="hljs bash">wget https://cf.10xgenomics.com/samples/cell/pbmc3k/pbmc3k_filtered_gene_bc_matrices.tar.gz<br>tar -zxvf pbmc3k_filtered_gene_bc_matrices.tar.gz<br></code></pre></td></tr></table></figure>
- <h2 id="读入数据">读入数据</h2>
- <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br><span class="line">5</span><br><span class="line">6</span><br><span class="line">7</span><br><span class="line">8</span><br><span class="line">9</span><br><span class="line">10</span><br><span class="line">11</span><br></pre></td><td class="code"><pre><code class="hljs R">pbmc <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>Read10X<span class="hljs-punctuation">(</span><span class="hljs-string">'~/data/filtered_gene_bc_matrices/hg19/'</span><span class="hljs-punctuation">)</span><br>sce <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>CreateSeuratObject<span class="hljs-punctuation">(</span>pbmc<span class="hljs-punctuation">,</span> project <span class="hljs-operator">=</span> <span class="hljs-string">'pbmc3k'</span><span class="hljs-punctuation">,</span><br> min.cells <span class="hljs-operator">=</span> <span class="hljs-number">3</span><span class="hljs-punctuation">,</span> min.features <span class="hljs-operator">=</span> <span class="hljs-number">200</span><span class="hljs-punctuation">)</span><br><span class="hljs-comment"># rm(pbmc);gc()</span><br><span class="hljs-comment"># 线粒体基因</span><br>sce<span class="hljs-punctuation">[[</span><span class="hljs-string">"percent.mt"</span><span class="hljs-punctuation">]</span><span class="hljs-punctuation">]</span> <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>PercentageFeatureSet<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> pattern <span class="hljs-operator">=</span> <span class="hljs-string">"^MT-"</span><span class="hljs-punctuation">)</span><br><span class="hljs-comment"># 核糖体基因</span><br>sce<span class="hljs-punctuation">[[</span><span class="hljs-string">"percent.ribo"</span><span class="hljs-punctuation">]</span><span class="hljs-punctuation">]</span> <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>PercentageFeatureSet<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> pattern <span class="hljs-operator">=</span> <span class="hljs-string">"^RP(S|L)"</span><span class="hljs-punctuation">)</span><br><span class="hljs-comment"># 血红蛋白基因 以 "HB" 开头,并且后面紧跟的字符不是 "P" </span><br>sce<span class="hljs-punctuation">[[</span><span class="hljs-string">"percent.hb"</span><span class="hljs-punctuation">]</span><span class="hljs-punctuation">]</span> <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>PercentageFeatureSet<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> pattern <span class="hljs-operator">=</span> <span class="hljs-string">"^HB[^(P)]"</span><span class="hljs-punctuation">)</span><br>sce<span class="hljs-operator">@</span>meta.data<br></code></pre></td></tr></table></figure>
- <h2 id="环境-RNA-校正">环境 RNA 校正</h2>
- <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br></pre></td><td class="code"><pre><code class="hljs R">decontX_results <span class="hljs-operator"><-</span> decontX<span class="hljs-operator">::</span>decontX<span class="hljs-punctuation">(</span>sce<span class="hljs-operator">@</span>assays<span class="hljs-operator">$</span>RNA<span class="hljs-operator">@</span>counts<span class="hljs-punctuation">)</span> <br>sce<span class="hljs-operator">$</span>Contamination <span class="hljs-operator">=</span> decontX_results<span class="hljs-operator">$</span>contamination<br>sce<span class="hljs-operator">@</span>meta.data<br></code></pre></td></tr></table></figure>
- <h2 id="计算细胞周期评分">计算细胞周期评分</h2>
- <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br><span class="line">5</span><br><span class="line">6</span><br><span class="line">7</span><br></pre></td><td class="code"><pre><code class="hljs R">sce <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>NormalizeData<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">)</span><br>g2m_genes <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>CaseMatch<span class="hljs-punctuation">(</span>search<span class="hljs-operator">=</span>Seurat<span class="hljs-operator">::</span>cc.genes<span class="hljs-operator">$</span>g2m.genes<span class="hljs-punctuation">,</span> <br> match<span class="hljs-operator">=</span>rownames<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br>s_genes <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>CaseMatch<span class="hljs-punctuation">(</span>search<span class="hljs-operator">=</span>Seurat<span class="hljs-operator">::</span>cc.genes<span class="hljs-operator">$</span>s.genes<span class="hljs-punctuation">,</span> <br> match<span class="hljs-operator">=</span>rownames<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br>sce <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>CellCycleScoring<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> g2m.features<span class="hljs-operator">=</span>g2m_genes<span class="hljs-punctuation">,</span> s.features<span class="hljs-operator">=</span>s_genes<span class="hljs-punctuation">)</span><br>sce<span class="hljs-operator">$</span>CC.Difference <span class="hljs-operator"><-</span> sce<span class="hljs-operator">$</span>S.Score <span class="hljs-operator">-</span> sce<span class="hljs-operator">$</span>G2M.Score<br></code></pre></td></tr></table></figure>
- <h2 id="双细胞过滤">双细胞过滤</h2>
- <ul>
- <li><a href="/shi-yong-SCTransform-biao-zhun-hua">更多细节</a></li>
- </ul>
- <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br></pre></td><td class="code"><pre><code class="hljs R">sce <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>SCTransform<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> vst.flavor <span class="hljs-operator">=</span> <span class="hljs-string">"v2"</span><span class="hljs-punctuation">,</span><br> vars.to.regress <span class="hljs-operator">=</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-string">"percent.mt"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"percent.ribo"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"percent.hb"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"Contamination"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"CC.Difference"</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><br> verbose <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">)</span><br></code></pre></td></tr></table></figure>
- <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br></pre></td><td class="code"><pre><code class="hljs R">sweep.res <span class="hljs-operator"><-</span> DoubletFinder<span class="hljs-operator">::</span>paramSweep_v3<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> PCs <span class="hljs-operator">=</span> <span class="hljs-number">1</span><span class="hljs-operator">:</span><span class="hljs-number">10</span><span class="hljs-punctuation">,</span> sct <span class="hljs-operator">=</span> <span class="hljs-built_in">T</span><span class="hljs-punctuation">,</span> num.cores<span class="hljs-operator">=</span><span class="hljs-number">8</span><span class="hljs-punctuation">)</span><br>sweep.stats <span class="hljs-operator"><-</span> DoubletFinder<span class="hljs-operator">::</span>summarizeSweep<span class="hljs-punctuation">(</span>sweep.res<span class="hljs-punctuation">,</span> GT <span class="hljs-operator">=</span> <span class="hljs-literal">FALSE</span><span class="hljs-punctuation">)</span><br>bcmvn <span class="hljs-operator"><-</span> DoubletFinder<span class="hljs-operator">::</span>find.pK<span class="hljs-punctuation">(</span>sweep.stats<span class="hljs-punctuation">)</span><br></code></pre></td></tr></table></figure>
- <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br><span class="line">5</span><br><span class="line">6</span><br><span class="line">7</span><br><span class="line">8</span><br></pre></td><td class="code"><pre><code class="hljs R"><span class="hljs-comment">## Assuming 1.6% doublet formation rate - tailor for your dataset</span><br>nExp_poi <span class="hljs-operator"><-</span> <span class="hljs-built_in">round</span><span class="hljs-punctuation">(</span><span class="hljs-number">0.016</span><span class="hljs-operator">*</span>nrow<span class="hljs-punctuation">(</span>sce<span class="hljs-operator">@</span>meta.data<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br>pK_bcmvn <span class="hljs-operator"><-</span> <span class="hljs-built_in">as.numeric</span><span class="hljs-punctuation">(</span><span class="hljs-built_in">as.character</span><span class="hljs-punctuation">(</span>bcmvn<span class="hljs-operator">$</span>pK<span class="hljs-punctuation">[</span>which.max<span class="hljs-punctuation">(</span>bcmvn<span class="hljs-operator">$</span>BCmetric<span class="hljs-punctuation">)</span><span class="hljs-punctuation">]</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br>sce <span class="hljs-operator"><-</span> DoubletFinder<span class="hljs-operator">::</span>doubletFinder_v3<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> PCs <span class="hljs-operator">=</span> <span class="hljs-number">1</span><span class="hljs-operator">:</span><span class="hljs-number">10</span><span class="hljs-punctuation">,</span> <br> pN <span class="hljs-operator">=</span> <span class="hljs-number">0.25</span><span class="hljs-punctuation">,</span> pK <span class="hljs-operator">=</span> pK_bcmvn<span class="hljs-punctuation">,</span> <br> nExp <span class="hljs-operator">=</span> nExp_poi<span class="hljs-punctuation">,</span> reuse.pANN <span class="hljs-operator">=</span> <span class="hljs-literal">FALSE</span><span class="hljs-punctuation">,</span> <br> sct <span class="hljs-operator">=</span> <span class="hljs-built_in">T</span><span class="hljs-punctuation">)</span><br>sce<span class="hljs-operator">@</span>meta.data<br></code></pre></td></tr></table></figure>
- <h2 id="可视化">可视化</h2>
- <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br><span class="line">5</span><br><span class="line">6</span><br><span class="line">7</span><br><span class="line">8</span><br><span class="line">9</span><br><span class="line">10</span><br><span class="line">11</span><br><span class="line">12</span><br><span class="line">13</span><br><span class="line">14</span><br><span class="line">15</span><br><span class="line">16</span><br><span class="line">17</span><br><span class="line">18</span><br></pre></td><td class="code"><pre><code class="hljs R">sce <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>RunPCA<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> assay<span class="hljs-operator">=</span><span class="hljs-string">"SCT"</span><span class="hljs-punctuation">,</span> verbose <span class="hljs-operator">=</span> <span class="hljs-literal">FALSE</span><span class="hljs-punctuation">)</span><br>Seurat<span class="hljs-operator">::</span>ElbowPlot<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> ndims <span class="hljs-operator">=</span> <span class="hljs-number">50</span><span class="hljs-punctuation">)</span><br>sce <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>RunUMAP<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> reduction <span class="hljs-operator">=</span> <span class="hljs-string">"pca"</span><span class="hljs-punctuation">,</span> <br> dims <span class="hljs-operator">=</span> <span class="hljs-number">1</span><span class="hljs-operator">:</span><span class="hljs-number">30</span><span class="hljs-punctuation">,</span> verbose <span class="hljs-operator">=</span> <span class="hljs-literal">FALSE</span><span class="hljs-punctuation">)</span><br>sce <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>FindNeighbors<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> reduction <span class="hljs-operator">=</span> <span class="hljs-string">"pca"</span><span class="hljs-punctuation">,</span> <br> dims <span class="hljs-operator">=</span> <span class="hljs-number">1</span><span class="hljs-operator">:</span><span class="hljs-number">30</span><span class="hljs-punctuation">,</span> verbose <span class="hljs-operator">=</span> <span class="hljs-literal">FALSE</span><span class="hljs-punctuation">)</span><br>options<span class="hljs-punctuation">(</span>repr.plot.width <span class="hljs-operator">=</span> <span class="hljs-number">8</span><span class="hljs-punctuation">,</span> repr.plot.height <span class="hljs-operator">=</span> <span class="hljs-number">12</span><span class="hljs-punctuation">)</span><br>FeaturePlot<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> <br> features <span class="hljs-operator">=</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-string">"percent.mt"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"percent.ribo"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"percent.hb"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"Contamination"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"CC.Difference"</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> <br> raster<span class="hljs-operator">=</span><span class="hljs-literal">FALSE</span> <span class="hljs-comment"># 细胞过多时候需要加这个参数</span><br> <span class="hljs-punctuation">)</span><br>options<span class="hljs-punctuation">(</span>repr.plot.width <span class="hljs-operator">=</span> <span class="hljs-number">6</span><span class="hljs-punctuation">,</span> repr.plot.height <span class="hljs-operator">=</span> <span class="hljs-number">6</span><span class="hljs-punctuation">)</span><br>DimPlot<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> reduction <span class="hljs-operator">=</span> <span class="hljs-string">"umap"</span><span class="hljs-punctuation">,</span> group.by <span class="hljs-operator">=</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-string">"DF.classifications_0.25_0.05_43"</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br>options<span class="hljs-punctuation">(</span>repr.plot.width <span class="hljs-operator">=</span> <span class="hljs-number">12</span><span class="hljs-punctuation">,</span> repr.plot.height <span class="hljs-operator">=</span> <span class="hljs-number">6</span><span class="hljs-punctuation">)</span><br>Seurat<span class="hljs-operator">::</span>VlnPlot<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> features <span class="hljs-operator">=</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-string">"nFeature_RNA"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"nCount_RNA"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"percent.mt"</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> ncol <span class="hljs-operator">=</span> <span class="hljs-number">3</span><span class="hljs-punctuation">)</span><br>plot1 <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>FeatureScatter<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> feature1 <span class="hljs-operator">=</span> <span class="hljs-string">"nCount_RNA"</span><span class="hljs-punctuation">,</span> feature2 <span class="hljs-operator">=</span> <span class="hljs-string">"percent.mt"</span><span class="hljs-punctuation">)</span><br>plot2 <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>FeatureScatter<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> feature1 <span class="hljs-operator">=</span> <span class="hljs-string">"nCount_RNA"</span><span class="hljs-punctuation">,</span> feature2 <span class="hljs-operator">=</span> <span class="hljs-string">"nFeature_RNA"</span><span class="hljs-punctuation">)</span><br>plot1 <span class="hljs-operator">+</span> plot2<br></code></pre></td></tr></table></figure>
- <h2 id="过滤低质量细胞">过滤低质量细胞</h2>
- <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br></pre></td><td class="code"><pre><code class="hljs R">sce <span class="hljs-operator"><-</span> subset<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> DF.classifications_0.25_0.04_43 <span class="hljs-operator">!=</span> <span class="hljs-string">'Doublet'</span><br> <span class="hljs-operator">&</span> Contamination <span class="hljs-operator"><</span> <span class="hljs-number">0.2</span><br><span class="hljs-punctuation">)</span><br></code></pre></td></tr></table></figure>
- <h2 id="查看分群走向">查看分群走向</h2>
- <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br><span class="line">5</span><br></pre></td><td class="code"><pre><code class="hljs R">sce <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>SCTransform<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> vst.flavor <span class="hljs-operator">=</span> <span class="hljs-string">"v2"</span><span class="hljs-punctuation">,</span><br> vars.to.regress <span class="hljs-operator">=</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-string">"percent.mt"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"percent.ribo"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"percent.hb"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"Contamination"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"CC.Difference"</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><br> verbose <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">)</span><br>sce <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>RunPCA<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> assay<span class="hljs-operator">=</span><span class="hljs-string">"SCT"</span><span class="hljs-punctuation">,</span> verbose <span class="hljs-operator">=</span> <span class="hljs-literal">FALSE</span><span class="hljs-punctuation">)</span><br>Seurat<span class="hljs-operator">::</span>ElbowPlot<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> ndims <span class="hljs-operator">=</span> <span class="hljs-number">50</span><span class="hljs-punctuation">)</span><br></code></pre></td></tr></table></figure>
- <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br></pre></td><td class="code"><pre><code class="hljs R">sce <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>RunUMAP<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> reduction <span class="hljs-operator">=</span> <span class="hljs-string">"pca"</span><span class="hljs-punctuation">,</span> <br> dims <span class="hljs-operator">=</span> <span class="hljs-number">1</span><span class="hljs-operator">:</span><span class="hljs-number">12</span><span class="hljs-punctuation">,</span> verbose <span class="hljs-operator">=</span> <span class="hljs-literal">FALSE</span><span class="hljs-punctuation">)</span><br>sce <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>FindNeighbors<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> reduction <span class="hljs-operator">=</span> <span class="hljs-string">"pca"</span><span class="hljs-punctuation">,</span> <br> dims <span class="hljs-operator">=</span> <span class="hljs-number">1</span><span class="hljs-operator">:</span><span class="hljs-number">12</span><span class="hljs-punctuation">,</span> verbose <span class="hljs-operator">=</span> <span class="hljs-literal">FALSE</span><span class="hljs-punctuation">)</span><br></code></pre></td></tr></table></figure>
- <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br><span class="line">5</span><br><span class="line">6</span><br><span class="line">7</span><br></pre></td><td class="code"><pre><code class="hljs R">sce <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>FindClusters<span class="hljs-punctuation">(</span><br> object <span class="hljs-operator">=</span> sce<span class="hljs-punctuation">,</span><br> resolution <span class="hljs-operator">=</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span>seq<span class="hljs-punctuation">(</span><span class="hljs-number">.1</span><span class="hljs-punctuation">,</span><span class="hljs-number">1.6</span><span class="hljs-punctuation">,</span><span class="hljs-number">.2</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span> <span class="hljs-comment">#起始粒度,结束粒度,间隔</span><br><span class="hljs-punctuation">)</span><br>options<span class="hljs-punctuation">(</span>repr.plot.width <span class="hljs-operator">=</span> <span class="hljs-number">12</span><span class="hljs-punctuation">,</span> repr.plot.height <span class="hljs-operator">=</span> <span class="hljs-number">12</span><span class="hljs-punctuation">)</span><br>require<span class="hljs-punctuation">(</span>clustree<span class="hljs-punctuation">)</span><br>clustree<span class="hljs-operator">::</span>clustree<span class="hljs-punctuation">(</span>sce<span class="hljs-operator">@</span>meta.data<span class="hljs-punctuation">,</span> prefix <span class="hljs-operator">=</span> <span class="hljs-string">"SCT_snn_res."</span><span class="hljs-punctuation">)</span><br></code></pre></td></tr></table></figure>
- <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br></pre></td><td class="code"><pre><code class="hljs R">options<span class="hljs-punctuation">(</span>repr.plot.width <span class="hljs-operator">=</span> <span class="hljs-number">6</span><span class="hljs-punctuation">,</span> repr.plot.height <span class="hljs-operator">=</span> <span class="hljs-number">6</span><span class="hljs-punctuation">)</span><br>DimPlot<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> reduction <span class="hljs-operator">=</span> <span class="hljs-string">"umap"</span><span class="hljs-punctuation">,</span> group.by <span class="hljs-operator">=</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-string">"SCT_snn_res.0.9"</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br></code></pre></td></tr></table></figure>
- <h2 id="可视化-2">可视化</h2>
- <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br><span class="line">5</span><br><span class="line">6</span><br><span class="line">7</span><br><span class="line">8</span><br><span class="line">9</span><br><span class="line">10</span><br><span class="line">11</span><br><span class="line">12</span><br><span class="line">13</span><br><span class="line">14</span><br></pre></td><td class="code"><pre><code class="hljs R">sub_celltype <span class="hljs-operator"><-</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-string">"0"</span><span class="hljs-punctuation">,</span><span class="hljs-string">"1"</span> <span class="hljs-punctuation">,</span><span class="hljs-string">"2"</span> <span class="hljs-punctuation">,</span><span class="hljs-string">"3"</span><span class="hljs-punctuation">)</span><br>sub.celltype_list <span class="hljs-operator"><-</span> sapply<span class="hljs-punctuation">(</span>sub_celltype<span class="hljs-punctuation">,</span><span class="hljs-keyword">function</span><span class="hljs-punctuation">(</span>slide<span class="hljs-punctuation">)</span><span class="hljs-punctuation">{</span><br> print<span class="hljs-punctuation">(</span>slide<span class="hljs-punctuation">)</span><br> sub.celltype <span class="hljs-operator"><-</span> subset<span class="hljs-punctuation">(</span>sce <span class="hljs-punctuation">,</span> `SCT_snn_res.0.1` <span class="hljs-operator">==</span> slide <span class="hljs-punctuation">)</span><br> sub.celltype <span class="hljs-operator"><-</span> Seurat<span class="hljs-operator">::</span>SCTransform<span class="hljs-punctuation">(</span>sub.celltype<span class="hljs-punctuation">,</span> vst.flavor <span class="hljs-operator">=</span> <span class="hljs-string">"v2"</span><span class="hljs-punctuation">,</span><br> vars.to.regress <span class="hljs-operator">=</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-string">"percent.mt"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"percent.ribo"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"percent.hb"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"Contamination"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"CC.Difference"</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><br> verbose <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">)</span><br> sub.celltype <span class="hljs-operator"><-</span> RunPCA<span class="hljs-punctuation">(</span>sub.celltype<span class="hljs-punctuation">,</span> npcs <span class="hljs-operator">=</span> <span class="hljs-number">20</span><span class="hljs-punctuation">)</span><br> sub.celltype <span class="hljs-operator"><-</span> FindNeighbors<span class="hljs-punctuation">(</span>sub.celltype<span class="hljs-punctuation">,</span> dims <span class="hljs-operator">=</span> <span class="hljs-number">1</span><span class="hljs-operator">:</span><span class="hljs-number">20</span><span class="hljs-punctuation">)</span><br> sub.celltype <span class="hljs-operator"><-</span> FindClusters<span class="hljs-punctuation">(</span>sub.celltype<span class="hljs-punctuation">,</span> resolution <span class="hljs-operator">=</span> <span class="hljs-number">0.9</span><span class="hljs-punctuation">)</span><br> sub.celltype <span class="hljs-operator"><-</span> RunUMAP<span class="hljs-punctuation">(</span>sub.celltype<span class="hljs-punctuation">,</span> dims <span class="hljs-operator">=</span> <span class="hljs-number">1</span><span class="hljs-operator">:</span><span class="hljs-number">20</span><span class="hljs-punctuation">)</span><br> <span class="hljs-built_in">return</span><span class="hljs-punctuation">(</span>sub.celltype<span class="hljs-punctuation">)</span><br><span class="hljs-punctuation">}</span><span class="hljs-punctuation">)</span> <br>sub.celltype_list<br></code></pre></td></tr></table></figure>
- <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br><span class="line">5</span><br><span class="line">6</span><br><span class="line">7</span><br><span class="line">8</span><br><span class="line">9</span><br><span class="line">10</span><br><span class="line">11</span><br><span class="line">12</span><br><span class="line">13</span><br><span class="line">14</span><br><span class="line">15</span><br><span class="line">16</span><br><span class="line">17</span><br><span class="line">18</span><br><span class="line">19</span><br><span class="line">20</span><br><span class="line">21</span><br><span class="line">22</span><br><span class="line">23</span><br><span class="line">24</span><br><span class="line">25</span><br><span class="line">26</span><br><span class="line">27</span><br><span class="line">28</span><br><span class="line">29</span><br><span class="line">30</span><br><span class="line">31</span><br><span class="line">32</span><br><span class="line">33</span><br><span class="line">34</span><br><span class="line">35</span><br><span class="line">36</span><br><span class="line">37</span><br><span class="line">38</span><br><span class="line">39</span><br><span class="line">40</span><br><span class="line">41</span><br><span class="line">42</span><br><span class="line">43</span><br><span class="line">44</span><br><span class="line">45</span><br><span class="line">46</span><br><span class="line">47</span><br><span class="line">48</span><br><span class="line">49</span><br><span class="line">50</span><br><span class="line">51</span><br><span class="line">52</span><br><span class="line">53</span><br><span class="line">54</span><br><span class="line">55</span><br><span class="line">56</span><br><span class="line">57</span><br><span class="line">58</span><br><span class="line">59</span><br><span class="line">60</span><br><span class="line">61</span><br><span class="line">62</span><br><span class="line">63</span><br><span class="line">64</span><br><span class="line">65</span><br></pre></td><td class="code"><pre><code class="hljs R">require<span class="hljs-punctuation">(</span>tidyverse<span class="hljs-punctuation">)</span><br>require<span class="hljs-punctuation">(</span>plot1cell<span class="hljs-punctuation">)</span><br><br>my36colors <span class="hljs-operator"><-</span><span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-string">'#E5D2DD'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#53A85F'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#F1BB72'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#F3B1A0'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#D6E7A3'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#57C3F3'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#476D87'</span><span class="hljs-punctuation">,</span><br> <span class="hljs-string">'#E95C59'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#E59CC4'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#AB3282'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#23452F'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#BD956A'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#8C549C'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#585658'</span><span class="hljs-punctuation">,</span><br> <span class="hljs-string">'#9FA3A8'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#E0D4CA'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#5F3D69'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#C5DEBA'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#58A4C3'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#E4C755'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#F7F398'</span><span class="hljs-punctuation">,</span><br> <span class="hljs-string">'#AA9A59'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#E63863'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#E39A35'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#C1E6F3'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#6778AE'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#91D0BE'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#B53E2B'</span><span class="hljs-punctuation">,</span><br> <span class="hljs-string">'#712820'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#DCC1DD'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#CCE0F5'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#CCC9E6'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#625D9E'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#68A180'</span><span class="hljs-punctuation">,</span> <span class="hljs-string">'#3A6963'</span><span class="hljs-punctuation">,</span><br> <span class="hljs-string">'#968175'</span><br><span class="hljs-punctuation">)</span><br><br>plot_circlize_change <span class="hljs-operator"><-</span> <span class="hljs-keyword">function</span> <span class="hljs-punctuation">(</span>data_plot<span class="hljs-punctuation">,</span> do.label <span class="hljs-operator">=</span> <span class="hljs-built_in">T</span><span class="hljs-punctuation">,</span> contour.levels <span class="hljs-operator">=</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-number">0.2</span><span class="hljs-punctuation">,</span> <span class="hljs-number">0.3</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> <br> pt.size <span class="hljs-operator">=</span> <span class="hljs-number">0.5</span><span class="hljs-punctuation">,</span> kde2d.n <span class="hljs-operator">=</span> <span class="hljs-number">1000</span><span class="hljs-punctuation">,</span> contour.nlevels <span class="hljs-operator">=</span> <span class="hljs-number">100</span><span class="hljs-punctuation">,</span> bg.color <span class="hljs-operator">=</span> <span class="hljs-string">"#F9F2E4"</span><span class="hljs-punctuation">,</span> <br> col.use <span class="hljs-operator">=</span> <span class="hljs-literal">NULL</span><span class="hljs-punctuation">,</span> label.cex <span class="hljs-operator">=</span> <span class="hljs-number">0.5</span><span class="hljs-punctuation">,</span> labels.cex <span class="hljs-operator">=</span> <span class="hljs-number">0.5</span><span class="hljs-punctuation">,</span> circos.cex <span class="hljs-operator">=</span> <span class="hljs-number">0.5</span> <span class="hljs-punctuation">,</span>repel <span class="hljs-operator">=</span> <span class="hljs-literal">FALSE</span><span class="hljs-punctuation">)</span> <br><span class="hljs-punctuation">{</span><br> centers <span class="hljs-operator"><-</span> data_plot <span class="hljs-operator">%>%</span> dplyr<span class="hljs-operator">::</span>group_by<span class="hljs-punctuation">(</span>Cluster<span class="hljs-punctuation">)</span> <span class="hljs-operator">%>%</span> summarise<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> median<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> x<span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> <br> y <span class="hljs-operator">=</span> median<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> y<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br> z <span class="hljs-operator"><-</span> MASS<span class="hljs-operator">::</span>kde2d<span class="hljs-punctuation">(</span>data_plot<span class="hljs-operator">$</span>x<span class="hljs-punctuation">,</span> data_plot<span class="hljs-operator">$</span>y<span class="hljs-punctuation">,</span> n <span class="hljs-operator">=</span> kde2d.n<span class="hljs-punctuation">)</span><br> celltypes <span class="hljs-operator"><-</span> <span class="hljs-built_in">names</span><span class="hljs-punctuation">(</span>table<span class="hljs-punctuation">(</span>data_plot<span class="hljs-operator">$</span>Cluster<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br> cell_colors <span class="hljs-operator"><-</span> <span class="hljs-punctuation">(</span>scales<span class="hljs-operator">::</span>hue_pal<span class="hljs-punctuation">(</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">(</span><span class="hljs-built_in">length</span><span class="hljs-punctuation">(</span>celltypes<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br> <span class="hljs-keyword">if</span> <span class="hljs-punctuation">(</span><span class="hljs-operator">!</span><span class="hljs-built_in">is.null</span><span class="hljs-punctuation">(</span>col.use<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span> <span class="hljs-punctuation">{</span><br> cell_colors <span class="hljs-operator">=</span> col.use<br> col_df <span class="hljs-operator"><-</span> data.frame<span class="hljs-punctuation">(</span>Cluster <span class="hljs-operator">=</span> celltypes<span class="hljs-punctuation">,</span> color2 <span class="hljs-operator">=</span> col.use<span class="hljs-punctuation">)</span><br> cells_order <span class="hljs-operator"><-</span> rownames<span class="hljs-punctuation">(</span>data_plot<span class="hljs-punctuation">)</span><br> data_plot <span class="hljs-operator"><-</span> merge<span class="hljs-punctuation">(</span>data_plot<span class="hljs-punctuation">,</span> col_df<span class="hljs-punctuation">,</span> by <span class="hljs-operator">=</span> <span class="hljs-string">"Cluster"</span><span class="hljs-punctuation">)</span><br> rownames<span class="hljs-punctuation">(</span>data_plot<span class="hljs-punctuation">)</span> <span class="hljs-operator"><-</span> data_plot<span class="hljs-operator">$</span>cells<br> data_plot <span class="hljs-operator"><-</span> data_plot<span class="hljs-punctuation">[</span>cells_order<span class="hljs-punctuation">,</span> <span class="hljs-punctuation">]</span><br> data_plot<span class="hljs-operator">$</span>Colors <span class="hljs-operator"><-</span> data_plot<span class="hljs-operator">$</span>color2<br> <span class="hljs-punctuation">}</span><br> circos.clear<span class="hljs-punctuation">(</span><span class="hljs-punctuation">)</span><br> par<span class="hljs-punctuation">(</span>bg <span class="hljs-operator">=</span> bg.color<span class="hljs-punctuation">)</span><br> circos.par<span class="hljs-punctuation">(</span>cell.padding <span class="hljs-operator">=</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-number">0</span><span class="hljs-punctuation">,</span> <span class="hljs-number">0</span><span class="hljs-punctuation">,</span> <span class="hljs-number">0</span><span class="hljs-punctuation">,</span> <span class="hljs-number">0</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> track.margin <span class="hljs-operator">=</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-number">0.01</span><span class="hljs-punctuation">,</span> <br> <span class="hljs-number">0</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> track.height <span class="hljs-operator">=</span> <span class="hljs-number">0.01</span><span class="hljs-punctuation">,</span> gap.degree <span class="hljs-operator">=</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-built_in">rep</span><span class="hljs-punctuation">(</span><span class="hljs-number">2</span><span class="hljs-punctuation">,</span> <span class="hljs-punctuation">(</span><span class="hljs-built_in">length</span><span class="hljs-punctuation">(</span>celltypes<span class="hljs-punctuation">)</span> <span class="hljs-operator">-</span> <br> <span class="hljs-number">1</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> <span class="hljs-number">12</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> points.overflow.warning <span class="hljs-operator">=</span> <span class="hljs-literal">FALSE</span><span class="hljs-punctuation">)</span><br> circos.initialize<span class="hljs-punctuation">(</span>sectors <span class="hljs-operator">=</span> data_plot<span class="hljs-operator">$</span>Cluster<span class="hljs-punctuation">,</span> x <span class="hljs-operator">=</span> data_plot<span class="hljs-operator">$</span>x_polar2<span class="hljs-punctuation">)</span><br> circos.track<span class="hljs-punctuation">(</span>data_plot<span class="hljs-operator">$</span>Cluster<span class="hljs-punctuation">,</span> data_plot<span class="hljs-operator">$</span>x_polar2<span class="hljs-punctuation">,</span> y <span class="hljs-operator">=</span> data_plot<span class="hljs-operator">$</span>dim2<span class="hljs-punctuation">,</span> <br> bg.border <span class="hljs-operator">=</span> <span class="hljs-literal">NA</span><span class="hljs-punctuation">,</span> panel.fun <span class="hljs-operator">=</span> <span class="hljs-keyword">function</span><span class="hljs-punctuation">(</span>x<span class="hljs-punctuation">,</span> y<span class="hljs-punctuation">)</span> <span class="hljs-punctuation">{</span><br> circos.text<span class="hljs-punctuation">(</span>CELL_META<span class="hljs-operator">$</span>xcenter<span class="hljs-punctuation">,</span> CELL_META<span class="hljs-operator">$</span>cell.ylim<span class="hljs-punctuation">[</span><span class="hljs-number">2</span><span class="hljs-punctuation">]</span> <span class="hljs-operator">+</span> <br> mm_y<span class="hljs-punctuation">(</span><span class="hljs-number">4</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> CELL_META<span class="hljs-operator">$</span>sector.index<span class="hljs-punctuation">,</span> cex <span class="hljs-operator">=</span> labels.cex<span class="hljs-punctuation">,</span><br> col <span class="hljs-operator">=</span> <span class="hljs-string">"black"</span><span class="hljs-punctuation">,</span> facing <span class="hljs-operator">=</span> <span class="hljs-string">"bending.inside"</span><span class="hljs-punctuation">,</span> niceFacing <span class="hljs-operator">=</span> <span class="hljs-built_in">T</span><span class="hljs-punctuation">)</span><br> <span class="hljs-comment">#circos.axis(labels.cex = 0.3, col = "black", labels.col = "black")</span><br> circos.axis<span class="hljs-punctuation">(</span>labels.cex <span class="hljs-operator">=</span> circos.cex<span class="hljs-punctuation">,</span> col <span class="hljs-operator">=</span> <span class="hljs-string">"black"</span><span class="hljs-punctuation">,</span> labels.col <span class="hljs-operator">=</span> <span class="hljs-string">"black"</span><span class="hljs-punctuation">)</span><br> <span class="hljs-punctuation">}</span><span class="hljs-punctuation">)</span><br> <span class="hljs-keyword">for</span> <span class="hljs-punctuation">(</span>i <span class="hljs-keyword">in</span> <span class="hljs-number">1</span><span class="hljs-operator">:</span><span class="hljs-built_in">length</span><span class="hljs-punctuation">(</span>celltypes<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span> <span class="hljs-punctuation">{</span><br> dd <span class="hljs-operator"><-</span> data_plot<span class="hljs-punctuation">[</span>data_plot<span class="hljs-operator">$</span>Cluster <span class="hljs-operator">==</span> celltypes<span class="hljs-punctuation">[</span>i<span class="hljs-punctuation">]</span><span class="hljs-punctuation">,</span> <span class="hljs-punctuation">]</span><br> circos.segments<span class="hljs-punctuation">(</span>x0 <span class="hljs-operator">=</span> <span class="hljs-built_in">min</span><span class="hljs-punctuation">(</span>dd<span class="hljs-operator">$</span>x_polar2<span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> y0 <span class="hljs-operator">=</span> <span class="hljs-number">0</span><span class="hljs-punctuation">,</span> x1 <span class="hljs-operator">=</span> <span class="hljs-built_in">max</span><span class="hljs-punctuation">(</span>dd<span class="hljs-operator">$</span>x_polar2<span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> <br> y1 <span class="hljs-operator">=</span> <span class="hljs-number">0</span><span class="hljs-punctuation">,</span> col <span class="hljs-operator">=</span> cell_colors<span class="hljs-punctuation">[</span>i<span class="hljs-punctuation">]</span><span class="hljs-punctuation">,</span> lwd <span class="hljs-operator">=</span> <span class="hljs-number">3</span><span class="hljs-punctuation">,</span> sector.index <span class="hljs-operator">=</span> celltypes<span class="hljs-punctuation">[</span>i<span class="hljs-punctuation">]</span><span class="hljs-punctuation">)</span><br> <span class="hljs-punctuation">}</span><br> text<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> <span class="hljs-number">1</span><span class="hljs-punctuation">,</span> y <span class="hljs-operator">=</span> <span class="hljs-number">0.1</span><span class="hljs-punctuation">,</span> labels <span class="hljs-operator">=</span> <span class="hljs-string">"Cluster"</span><span class="hljs-punctuation">,</span> cex <span class="hljs-operator">=</span> <span class="hljs-number">0.4</span><span class="hljs-punctuation">,</span> col <span class="hljs-operator">=</span> <span class="hljs-string">"black"</span><span class="hljs-punctuation">,</span> <br> srt <span class="hljs-operator">=</span> <span class="hljs-operator">-</span><span class="hljs-number">90</span><span class="hljs-punctuation">)</span><br> points<span class="hljs-punctuation">(</span>data_plot<span class="hljs-operator">$</span>x<span class="hljs-punctuation">,</span> data_plot<span class="hljs-operator">$</span>y<span class="hljs-punctuation">,</span> pch <span class="hljs-operator">=</span> <span class="hljs-number">19</span><span class="hljs-punctuation">,</span> col <span class="hljs-operator">=</span> alpha<span class="hljs-punctuation">(</span>data_plot<span class="hljs-operator">$</span>Colors<span class="hljs-punctuation">,</span> <br> <span class="hljs-number">0.9</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> cex <span class="hljs-operator">=</span> pt.size<span class="hljs-punctuation">)</span><br> contour<span class="hljs-punctuation">(</span>z<span class="hljs-punctuation">,</span> drawlabels <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">,</span> nlevels <span class="hljs-operator">=</span> <span class="hljs-number">100</span><span class="hljs-punctuation">,</span> levels <span class="hljs-operator">=</span> contour.levels<span class="hljs-punctuation">,</span> <br> col <span class="hljs-operator">=</span> <span class="hljs-string">"#ae9c76"</span><span class="hljs-punctuation">,</span> add <span class="hljs-operator">=</span> <span class="hljs-literal">TRUE</span><span class="hljs-punctuation">)</span><br> <span class="hljs-keyword">if</span> <span class="hljs-punctuation">(</span>do.label<span class="hljs-punctuation">)</span> <span class="hljs-punctuation">{</span><br> <span class="hljs-keyword">if</span> <span class="hljs-punctuation">(</span>repel<span class="hljs-punctuation">)</span> <span class="hljs-punctuation">{</span><br> textplot<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> centers<span class="hljs-operator">$</span>x<span class="hljs-punctuation">,</span> y <span class="hljs-operator">=</span> centers<span class="hljs-operator">$</span>y<span class="hljs-punctuation">,</span> words <span class="hljs-operator">=</span> centers<span class="hljs-operator">$</span>Cluster<span class="hljs-punctuation">,</span> <br> cex <span class="hljs-operator">=</span> label.cex<span class="hljs-punctuation">,</span> new <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">,</span> show.lines <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">)</span><br> <span class="hljs-punctuation">}</span><br> <span class="hljs-keyword">else</span> <span class="hljs-punctuation">{</span><br> text<span class="hljs-punctuation">(</span>centers<span class="hljs-operator">$</span>x<span class="hljs-punctuation">,</span> centers<span class="hljs-operator">$</span>y<span class="hljs-punctuation">,</span> labels <span class="hljs-operator">=</span> centers<span class="hljs-operator">$</span>Cluster<span class="hljs-punctuation">,</span> <br> cex <span class="hljs-operator">=</span> label.cex<span class="hljs-punctuation">,</span> col <span class="hljs-operator">=</span> <span class="hljs-string">"black"</span><span class="hljs-punctuation">)</span><br> <span class="hljs-punctuation">}</span><br> <span class="hljs-punctuation">}</span><br><span class="hljs-punctuation">}</span><br></code></pre></td></tr></table></figure>
- <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br><span class="line">5</span><br><span class="line">6</span><br><span class="line">7</span><br><span class="line">8</span><br><span class="line">9</span><br><span class="line">10</span><br><span class="line">11</span><br><span class="line">12</span><br><span class="line">13</span><br><span class="line">14</span><br><span class="line">15</span><br><span class="line">16</span><br><span class="line">17</span><br><span class="line">18</span><br><span class="line">19</span><br><span class="line">20</span><br><span class="line">21</span><br><span class="line">22</span><br><span class="line">23</span><br><span class="line">24</span><br><span class="line">25</span><br><span class="line">26</span><br><span class="line">27</span><br><span class="line">28</span><br><span class="line">29</span><br><span class="line">30</span><br><span class="line">31</span><br><span class="line">32</span><br><span class="line">33</span><br><span class="line">34</span><br><span class="line">35</span><br><span class="line">36</span><br><span class="line">37</span><br><span class="line">38</span><br><span class="line">39</span><br><span class="line">40</span><br><span class="line">41</span><br><span class="line">42</span><br><span class="line">43</span><br><span class="line">44</span><br><span class="line">45</span><br><span class="line">46</span><br><span class="line">47</span><br><span class="line">48</span><br><span class="line">49</span><br><span class="line">50</span><br><span class="line">51</span><br><span class="line">52</span><br><span class="line">53</span><br><span class="line">54</span><br><span class="line">55</span><br><span class="line">56</span><br><span class="line">57</span><br><span class="line">58</span><br><span class="line">59</span><br><span class="line">60</span><br><span class="line">61</span><br><span class="line">62</span><br><span class="line">63</span><br><span class="line">64</span><br><span class="line">65</span><br><span class="line">66</span><br><span class="line">67</span><br><span class="line">68</span><br><span class="line">69</span><br><span class="line">70</span><br><span class="line">71</span><br><span class="line">72</span><br><span class="line">73</span><br><span class="line">74</span><br><span class="line">75</span><br><span class="line">76</span><br><span class="line">77</span><br><span class="line">78</span><br><span class="line">79</span><br><span class="line">80</span><br><span class="line">81</span><br><span class="line">82</span><br><span class="line">83</span><br><span class="line">84</span><br><span class="line">85</span><br><span class="line">86</span><br><span class="line">87</span><br><span class="line">88</span><br><span class="line">89</span><br><span class="line">90</span><br><span class="line">91</span><br><span class="line">92</span><br><span class="line">93</span><br><span class="line">94</span><br><span class="line">95</span><br><span class="line">96</span><br><span class="line">97</span><br><span class="line">98</span><br><span class="line">99</span><br><span class="line">100</span><br><span class="line">101</span><br><span class="line">102</span><br><span class="line">103</span><br><span class="line">104</span><br><span class="line">105</span><br><span class="line">106</span><br><span class="line">107</span><br><span class="line">108</span><br><span class="line">109</span><br><span class="line">110</span><br><span class="line">111</span><br><span class="line">112</span><br><span class="line">113</span><br><span class="line">114</span><br><span class="line">115</span><br><span class="line">116</span><br><span class="line">117</span><br><span class="line">118</span><br><span class="line">119</span><br></pre></td><td class="code"><pre><code class="hljs R"><span class="hljs-comment">###Prepare data for ploting 准备圈图数据</span><br>circ_data <span class="hljs-operator"><-</span> plot1cell<span class="hljs-operator">::</span>prepare_circlize_data<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> scale <span class="hljs-operator">=</span> <span class="hljs-number">0.7</span><span class="hljs-punctuation">)</span> <span class="hljs-comment"># 画出框了就调小 scale</span><br>set.seed<span class="hljs-punctuation">(</span><span class="hljs-number">1234</span><span class="hljs-punctuation">)</span><br><br><span class="hljs-comment"># 设置细胞分群信息的颜色</span><br>cluster_colors <span class="hljs-operator"><-</span> circlize<span class="hljs-operator">::</span>rand_color<span class="hljs-punctuation">(</span><span class="hljs-built_in">length</span><span class="hljs-punctuation">(</span>levels<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br>group_colors <span class="hljs-operator"><-</span> circlize<span class="hljs-operator">::</span>rand_color<span class="hljs-punctuation">(</span><span class="hljs-built_in">length</span><span class="hljs-punctuation">(</span><span class="hljs-built_in">names</span><span class="hljs-punctuation">(</span>table<span class="hljs-punctuation">(</span>sce<span class="hljs-operator">$</span>`SCT_snn_res.0.1`<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br>rep_colors <span class="hljs-operator"><-</span> circlize<span class="hljs-operator">::</span>rand_color<span class="hljs-punctuation">(</span><span class="hljs-built_in">length</span><span class="hljs-punctuation">(</span><span class="hljs-built_in">names</span><span class="hljs-punctuation">(</span>table<span class="hljs-punctuation">(</span>sce<span class="hljs-operator">$</span>orig.ident<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br><br><span class="hljs-comment"># 绘制细胞分群圈图</span><br><span class="hljs-punctuation">{</span>pdf<span class="hljs-punctuation">(</span>file <span class="hljs-operator">=</span> <span class="hljs-string">'umap_circlize.plot.pdf'</span><span class="hljs-punctuation">,</span> width <span class="hljs-operator">=</span> <span class="hljs-number">9</span><span class="hljs-punctuation">,</span> height <span class="hljs-operator">=</span> <span class="hljs-number">6</span><span class="hljs-punctuation">)</span> <span class="hljs-comment"># 导出PDF开始</span><br><br>options<span class="hljs-punctuation">(</span>repr.plot.width <span class="hljs-operator">=</span> <span class="hljs-number">12</span><span class="hljs-punctuation">,</span> repr.plot.height <span class="hljs-operator">=</span> <span class="hljs-number">8</span><span class="hljs-punctuation">)</span><br>plot_circlize_change<span class="hljs-punctuation">(</span>circ_data<span class="hljs-punctuation">,</span>do.label <span class="hljs-operator">=</span> <span class="hljs-built_in">T</span><span class="hljs-punctuation">,</span> pt.size <span class="hljs-operator">=</span> <span class="hljs-number">0.01</span><span class="hljs-punctuation">,</span> <br> col.use <span class="hljs-operator">=</span> cluster_colors <span class="hljs-punctuation">,</span><br> bg.color <span class="hljs-operator">=</span> <span class="hljs-string">'#F9F2E5'</span><span class="hljs-punctuation">,</span> <br> kde2d.n <span class="hljs-operator">=</span> <span class="hljs-number">1000</span><span class="hljs-punctuation">,</span> <br> repel <span class="hljs-operator">=</span> <span class="hljs-built_in">T</span><span class="hljs-punctuation">,</span> <br> labels.cex <span class="hljs-operator">=</span> <span class="hljs-number">1</span><span class="hljs-punctuation">,</span> <br> circos.cex <span class="hljs-operator">=</span> <span class="hljs-number">0.5</span><span class="hljs-punctuation">,</span><br> label.cex <span class="hljs-operator">=</span> <span class="hljs-number">1</span><span class="hljs-punctuation">)</span><br> <br>plot1cell<span class="hljs-operator">::</span>add_track<span class="hljs-punctuation">(</span>circ_data<span class="hljs-punctuation">,</span> <br> group <span class="hljs-operator">=</span> <span class="hljs-string">"SCT_snn_res.0.1"</span><span class="hljs-punctuation">,</span> <br> colors <span class="hljs-operator">=</span> group_colors<span class="hljs-punctuation">,</span> track_num <span class="hljs-operator">=</span> <span class="hljs-number">2</span><span class="hljs-punctuation">)</span> <span class="hljs-comment">## can change it to one of the columns in the meta data of your seurat object</span><br>plot1cell<span class="hljs-operator">::</span>add_track<span class="hljs-punctuation">(</span>circ_data<span class="hljs-punctuation">,</span> <br> group <span class="hljs-operator">=</span> <span class="hljs-string">"orig.ident"</span><span class="hljs-punctuation">,</span><br> colors <span class="hljs-operator">=</span> rep_colors<span class="hljs-punctuation">,</span> track_num <span class="hljs-operator">=</span> <span class="hljs-number">3</span><span class="hljs-punctuation">)</span><br><br><span class="hljs-comment"># 左下</span><br>sub_index <span class="hljs-operator">=</span> <span class="hljs-number">1</span><br>subcolors <span class="hljs-operator"><-</span> my36colors<span class="hljs-punctuation">[</span><span class="hljs-number">1</span><span class="hljs-operator">:</span>nlevels<span class="hljs-punctuation">(</span>sub.celltype_list<span class="hljs-punctuation">[[</span>sub_index<span class="hljs-punctuation">]</span><span class="hljs-punctuation">]</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">]</span><br>sub_1_meta<span class="hljs-operator"><-</span>get_metadata<span class="hljs-punctuation">(</span>sub.celltype_list<span class="hljs-punctuation">[[</span>sub_index<span class="hljs-punctuation">]</span><span class="hljs-punctuation">]</span><span class="hljs-punctuation">,</span> color <span class="hljs-operator">=</span> subcolors<span class="hljs-punctuation">)</span><br>sub_1_meta <span class="hljs-operator">%>%</span><br> dplyr<span class="hljs-operator">::</span>group_by<span class="hljs-punctuation">(</span>seurat_clusters<span class="hljs-punctuation">)</span> <span class="hljs-operator">%>%</span><br> summarize<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> median<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> x<span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span>y <span class="hljs-operator">=</span> median<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> y<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span> <span class="hljs-operator">-></span> centers_1<br><br>points<span class="hljs-punctuation">(</span>sub_1_meta<span class="hljs-operator">$</span>x<span class="hljs-operator">*</span><span class="hljs-number">0.32</span><span class="hljs-operator">-</span><span class="hljs-number">1.2</span><span class="hljs-punctuation">,</span>sub_1_meta<span class="hljs-operator">$</span>y<span class="hljs-operator">*</span><span class="hljs-number">0.32</span><span class="hljs-operator">-</span><span class="hljs-number">0.73</span><span class="hljs-punctuation">,</span> pch <span class="hljs-operator">=</span> <span class="hljs-number">19</span><span class="hljs-punctuation">,</span> col <span class="hljs-operator">=</span> alpha<span class="hljs-punctuation">(</span>sub_1_meta<span class="hljs-operator">$</span>Colors<span class="hljs-punctuation">,</span><span class="hljs-number">0.9</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> cex <span class="hljs-operator">=</span> <span class="hljs-number">0.1</span><span class="hljs-punctuation">)</span>;<br>text<span class="hljs-punctuation">(</span>centers_1<span class="hljs-operator">$</span>x<span class="hljs-operator">*</span><span class="hljs-number">0.32</span><span class="hljs-operator">-</span><span class="hljs-number">1.2</span><span class="hljs-punctuation">,</span>centers_1<span class="hljs-operator">$</span>y<span class="hljs-operator">*</span><span class="hljs-number">0.32</span><span class="hljs-operator">-</span><span class="hljs-number">0.73</span><span class="hljs-punctuation">,</span> labels<span class="hljs-operator">=</span>centers_1<span class="hljs-operator">$</span>seurat_clusters<span class="hljs-punctuation">,</span> cex <span class="hljs-operator">=</span> <span class="hljs-number">0.6</span><span class="hljs-punctuation">,</span> col <span class="hljs-operator">=</span> <span class="hljs-string">'black'</span><span class="hljs-punctuation">)</span><br><br><span class="hljs-comment"># 右上</span><br>sub_index <span class="hljs-operator">=</span> <span class="hljs-number">2</span><br>subcolors <span class="hljs-operator"><-</span> my36colors<span class="hljs-punctuation">[</span><span class="hljs-number">1</span><span class="hljs-operator">:</span>nlevels<span class="hljs-punctuation">(</span>sub.celltype_list<span class="hljs-punctuation">[[</span>sub_index<span class="hljs-punctuation">]</span><span class="hljs-punctuation">]</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">]</span><br>sub_1_meta<span class="hljs-operator"><-</span>get_metadata<span class="hljs-punctuation">(</span>sub.celltype_list<span class="hljs-punctuation">[[</span>sub_index<span class="hljs-punctuation">]</span><span class="hljs-punctuation">]</span><span class="hljs-punctuation">,</span> color <span class="hljs-operator">=</span> subcolors<span class="hljs-punctuation">)</span><br>sub_1_meta <span class="hljs-operator">%>%</span><br> dplyr<span class="hljs-operator">::</span>group_by<span class="hljs-punctuation">(</span>seurat_clusters<span class="hljs-punctuation">)</span> <span class="hljs-operator">%>%</span><br> summarize<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> median<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> x<span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span>y <span class="hljs-operator">=</span> median<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> y<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span> <span class="hljs-operator">-></span> centers_1<br><br>points<span class="hljs-punctuation">(</span>sub_1_meta<span class="hljs-operator">$</span>x<span class="hljs-operator">*</span><span class="hljs-number">0.32</span><span class="hljs-operator">+</span><span class="hljs-number">1.2</span><span class="hljs-punctuation">,</span>sub_1_meta<span class="hljs-operator">$</span>y<span class="hljs-operator">*</span><span class="hljs-number">0.32</span><span class="hljs-operator">+</span><span class="hljs-number">0.73</span><span class="hljs-punctuation">,</span> pch <span class="hljs-operator">=</span> <span class="hljs-number">19</span><span class="hljs-punctuation">,</span> col <span class="hljs-operator">=</span> alpha<span class="hljs-punctuation">(</span>sub_1_meta<span class="hljs-operator">$</span>Colors<span class="hljs-punctuation">,</span><span class="hljs-number">0.9</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> cex <span class="hljs-operator">=</span> <span class="hljs-number">0.1</span><span class="hljs-punctuation">)</span>;<br>text<span class="hljs-punctuation">(</span>centers_1<span class="hljs-operator">$</span>x<span class="hljs-operator">*</span><span class="hljs-number">0.32</span><span class="hljs-operator">+</span><span class="hljs-number">1.2</span><span class="hljs-punctuation">,</span>centers_1<span class="hljs-operator">$</span>y<span class="hljs-operator">*</span><span class="hljs-number">0.32</span><span class="hljs-operator">+</span><span class="hljs-number">0.73</span><span class="hljs-punctuation">,</span> labels<span class="hljs-operator">=</span>centers_1<span class="hljs-operator">$</span>seurat_clusters<span class="hljs-punctuation">,</span> cex <span class="hljs-operator">=</span> <span class="hljs-number">0.6</span><span class="hljs-punctuation">,</span> col <span class="hljs-operator">=</span> <span class="hljs-string">'black'</span><span class="hljs-punctuation">)</span><br><br><span class="hljs-comment"># 左上</span><br>sub_index <span class="hljs-operator">=</span> <span class="hljs-number">3</span><br>subcolors <span class="hljs-operator"><-</span> my36colors<span class="hljs-punctuation">[</span><span class="hljs-number">1</span><span class="hljs-operator">:</span>nlevels<span class="hljs-punctuation">(</span>sub.celltype_list<span class="hljs-punctuation">[[</span>sub_index<span class="hljs-punctuation">]</span><span class="hljs-punctuation">]</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">]</span><br>sub_1_meta<span class="hljs-operator"><-</span>get_metadata<span class="hljs-punctuation">(</span>sub.celltype_list<span class="hljs-punctuation">[[</span>sub_index<span class="hljs-punctuation">]</span><span class="hljs-punctuation">]</span><span class="hljs-punctuation">,</span> color <span class="hljs-operator">=</span> subcolors<span class="hljs-punctuation">)</span><br>sub_1_meta <span class="hljs-operator">%>%</span><br> dplyr<span class="hljs-operator">::</span>group_by<span class="hljs-punctuation">(</span>seurat_clusters<span class="hljs-punctuation">)</span> <span class="hljs-operator">%>%</span><br> summarize<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> median<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> x<span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span>y <span class="hljs-operator">=</span> median<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> y<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span> <span class="hljs-operator">-></span> centers_1<br><br>points<span class="hljs-punctuation">(</span>sub_1_meta<span class="hljs-operator">$</span>x<span class="hljs-operator">*</span><span class="hljs-number">0.32</span><span class="hljs-operator">-</span><span class="hljs-number">1.2</span><span class="hljs-punctuation">,</span>sub_1_meta<span class="hljs-operator">$</span>y<span class="hljs-operator">*</span><span class="hljs-number">0.32</span><span class="hljs-operator">+</span><span class="hljs-number">0.73</span><span class="hljs-punctuation">,</span> pch <span class="hljs-operator">=</span> <span class="hljs-number">19</span><span class="hljs-punctuation">,</span> col <span class="hljs-operator">=</span> alpha<span class="hljs-punctuation">(</span>sub_1_meta<span class="hljs-operator">$</span>Colors<span class="hljs-punctuation">,</span><span class="hljs-number">0.9</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> cex <span class="hljs-operator">=</span> <span class="hljs-number">0.1</span><span class="hljs-punctuation">)</span>;<br>text<span class="hljs-punctuation">(</span>centers_1<span class="hljs-operator">$</span>x<span class="hljs-operator">*</span><span class="hljs-number">0.32</span><span class="hljs-operator">-</span><span class="hljs-number">1.2</span><span class="hljs-punctuation">,</span>centers_1<span class="hljs-operator">$</span>y<span class="hljs-operator">*</span><span class="hljs-number">0.32</span><span class="hljs-operator">+</span><span class="hljs-number">0.73</span><span class="hljs-punctuation">,</span> labels<span class="hljs-operator">=</span>centers_1<span class="hljs-operator">$</span>seurat_clusters<span class="hljs-punctuation">,</span> cex <span class="hljs-operator">=</span> <span class="hljs-number">0.6</span><span class="hljs-punctuation">,</span> col <span class="hljs-operator">=</span> <span class="hljs-string">'black'</span><span class="hljs-punctuation">)</span><br><br><span class="hljs-comment"># 右下</span><br>sub_index <span class="hljs-operator">=</span> <span class="hljs-number">4</span><br>subcolors <span class="hljs-operator"><-</span> my36colors<span class="hljs-punctuation">[</span><span class="hljs-number">1</span><span class="hljs-operator">:</span>nlevels<span class="hljs-punctuation">(</span>sub.celltype_list<span class="hljs-punctuation">[[</span>sub_index<span class="hljs-punctuation">]</span><span class="hljs-punctuation">]</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">]</span><br>sub_1_meta<span class="hljs-operator"><-</span>get_metadata<span class="hljs-punctuation">(</span>sub.celltype_list<span class="hljs-punctuation">[[</span>sub_index<span class="hljs-punctuation">]</span><span class="hljs-punctuation">]</span><span class="hljs-punctuation">,</span> color <span class="hljs-operator">=</span> subcolors<span class="hljs-punctuation">)</span><br>sub_1_meta <span class="hljs-operator">%>%</span><br> dplyr<span class="hljs-operator">::</span>group_by<span class="hljs-punctuation">(</span>seurat_clusters<span class="hljs-punctuation">)</span> <span class="hljs-operator">%>%</span><br> summarize<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> median<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> x<span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span>y <span class="hljs-operator">=</span> median<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> y<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span> <span class="hljs-operator">-></span> centers_1<br><br>points<span class="hljs-punctuation">(</span>sub_1_meta<span class="hljs-operator">$</span>x<span class="hljs-operator">*</span><span class="hljs-number">0.32</span><span class="hljs-operator">+</span><span class="hljs-number">1.2</span><span class="hljs-punctuation">,</span>sub_1_meta<span class="hljs-operator">$</span>y<span class="hljs-operator">*</span><span class="hljs-number">0.32</span><span class="hljs-operator">-</span><span class="hljs-number">0.73</span><span class="hljs-punctuation">,</span> pch <span class="hljs-operator">=</span> <span class="hljs-number">19</span><span class="hljs-punctuation">,</span> col <span class="hljs-operator">=</span> alpha<span class="hljs-punctuation">(</span>sub_1_meta<span class="hljs-operator">$</span>Colors<span class="hljs-punctuation">,</span><span class="hljs-number">0.9</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> cex <span class="hljs-operator">=</span> <span class="hljs-number">0.1</span><span class="hljs-punctuation">)</span>;<br>text<span class="hljs-punctuation">(</span>centers_1<span class="hljs-operator">$</span>x<span class="hljs-operator">*</span><span class="hljs-number">0.32</span><span class="hljs-operator">+</span><span class="hljs-number">1.2</span><span class="hljs-punctuation">,</span>centers_1<span class="hljs-operator">$</span>y<span class="hljs-operator">*</span><span class="hljs-number">0.32</span><span class="hljs-operator">-</span><span class="hljs-number">0.73</span><span class="hljs-punctuation">,</span> labels<span class="hljs-operator">=</span>centers_1<span class="hljs-operator">$</span>seurat_clusters<span class="hljs-punctuation">,</span> cex <span class="hljs-operator">=</span> <span class="hljs-number">0.6</span><span class="hljs-punctuation">,</span> col <span class="hljs-operator">=</span> <span class="hljs-string">'black'</span><span class="hljs-punctuation">)</span><br><br><br>title_text <span class="hljs-operator"><-</span> <span class="hljs-keyword">function</span><span class="hljs-punctuation">(</span>x0<span class="hljs-punctuation">,</span> y0<span class="hljs-punctuation">,</span> x1<span class="hljs-punctuation">,</span> y1<span class="hljs-punctuation">,</span> text<span class="hljs-punctuation">,</span> rectArgs <span class="hljs-operator">=</span> <span class="hljs-literal">NULL</span><span class="hljs-punctuation">,</span> textArgs <span class="hljs-operator">=</span> <span class="hljs-literal">NULL</span><span class="hljs-punctuation">)</span> <span class="hljs-punctuation">{</span><br> center <span class="hljs-operator"><-</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span>mean<span class="hljs-punctuation">(</span><span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span>x0<span class="hljs-punctuation">,</span> x1<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> mean<span class="hljs-punctuation">(</span><span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span>y0<span class="hljs-punctuation">,</span> y1<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br> do.call<span class="hljs-punctuation">(</span><span class="hljs-string">'rect'</span><span class="hljs-punctuation">,</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-built_in">list</span><span class="hljs-punctuation">(</span>xleft <span class="hljs-operator">=</span> x0<span class="hljs-punctuation">,</span> ybottom <span class="hljs-operator">=</span> y0<span class="hljs-punctuation">,</span> xright <span class="hljs-operator">=</span> x1<span class="hljs-punctuation">,</span> ytop <span class="hljs-operator">=</span> y1<span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> rectArgs<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br> do.call<span class="hljs-punctuation">(</span><span class="hljs-string">'text'</span><span class="hljs-punctuation">,</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-built_in">list</span><span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> center<span class="hljs-punctuation">[</span><span class="hljs-number">1</span><span class="hljs-punctuation">]</span><span class="hljs-punctuation">,</span> y <span class="hljs-operator">=</span> center<span class="hljs-punctuation">[</span><span class="hljs-number">2</span><span class="hljs-punctuation">]</span><span class="hljs-punctuation">,</span> labels <span class="hljs-operator">=</span> text<span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> textArgs<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br><span class="hljs-punctuation">}</span><br><br><span class="hljs-comment"># 四周的title </span><br>title_text<span class="hljs-punctuation">(</span>x0 <span class="hljs-operator">=</span> <span class="hljs-operator">-</span><span class="hljs-number">1.35</span><span class="hljs-punctuation">,</span> x1 <span class="hljs-operator">=</span> <span class="hljs-operator">-</span><span class="hljs-number">1.05</span><span class="hljs-punctuation">,</span> y0 <span class="hljs-operator">=</span> <span class="hljs-operator">-</span><span class="hljs-number">1.06</span><span class="hljs-punctuation">,</span> y1<span class="hljs-operator">=</span><span class="hljs-operator">-</span><span class="hljs-number">1</span><span class="hljs-punctuation">,</span> text <span class="hljs-operator">=</span> <span class="hljs-string">'type 1 cells'</span><span class="hljs-punctuation">,</span><br> rectArgs <span class="hljs-operator">=</span> <span class="hljs-built_in">list</span><span class="hljs-punctuation">(</span>border<span class="hljs-operator">=</span><span class="hljs-string">'#F9F2E4'</span><span class="hljs-punctuation">,</span>lwd<span class="hljs-operator">=</span><span class="hljs-number">0.5</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><br> textArgs <span class="hljs-operator">=</span> <span class="hljs-built_in">list</span><span class="hljs-punctuation">(</span>col<span class="hljs-operator">=</span><span class="hljs-string">'black'</span><span class="hljs-punctuation">,</span>cex <span class="hljs-operator">=</span> <span class="hljs-number">1</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br><br>title_text<span class="hljs-punctuation">(</span>x0 <span class="hljs-operator">=</span> <span class="hljs-number">1.05</span><span class="hljs-punctuation">,</span> x1 <span class="hljs-operator">=</span> <span class="hljs-number">1.35</span><span class="hljs-punctuation">,</span> y0 <span class="hljs-operator">=</span> <span class="hljs-operator">-</span><span class="hljs-number">1.06</span><span class="hljs-punctuation">,</span> y1<span class="hljs-operator">=</span><span class="hljs-operator">-</span><span class="hljs-number">1</span><span class="hljs-punctuation">,</span> text <span class="hljs-operator">=</span> <span class="hljs-string">'type 2 cells'</span><span class="hljs-punctuation">,</span><br> rectArgs <span class="hljs-operator">=</span> <span class="hljs-built_in">list</span><span class="hljs-punctuation">(</span>border<span class="hljs-operator">=</span><span class="hljs-string">'#F9F2E4'</span><span class="hljs-punctuation">,</span>lwd<span class="hljs-operator">=</span><span class="hljs-number">0.5</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><br> textArgs <span class="hljs-operator">=</span> <span class="hljs-built_in">list</span><span class="hljs-punctuation">(</span>col<span class="hljs-operator">=</span><span class="hljs-string">'black'</span><span class="hljs-punctuation">,</span>cex <span class="hljs-operator">=</span> <span class="hljs-number">1</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br><br>title_text<span class="hljs-punctuation">(</span>x0 <span class="hljs-operator">=</span> <span class="hljs-operator">-</span><span class="hljs-number">1.35</span><span class="hljs-punctuation">,</span> x1 <span class="hljs-operator">=</span> <span class="hljs-operator">-</span><span class="hljs-number">1.05</span><span class="hljs-punctuation">,</span> y0 <span class="hljs-operator">=</span> <span class="hljs-number">1.06</span><span class="hljs-punctuation">,</span> y1<span class="hljs-operator">=</span><span class="hljs-number">1</span><span class="hljs-punctuation">,</span> text <span class="hljs-operator">=</span> <span class="hljs-string">'type 3 cells'</span><span class="hljs-punctuation">,</span><br> rectArgs <span class="hljs-operator">=</span> <span class="hljs-built_in">list</span><span class="hljs-punctuation">(</span>border<span class="hljs-operator">=</span><span class="hljs-string">'#F9F2E4'</span><span class="hljs-punctuation">,</span>lwd<span class="hljs-operator">=</span><span class="hljs-number">0.5</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><br> textArgs <span class="hljs-operator">=</span> <span class="hljs-built_in">list</span><span class="hljs-punctuation">(</span>col<span class="hljs-operator">=</span><span class="hljs-string">'black'</span><span class="hljs-punctuation">,</span>cex <span class="hljs-operator">=</span> <span class="hljs-number">1</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br><br>title_text<span class="hljs-punctuation">(</span>x0 <span class="hljs-operator">=</span> <span class="hljs-number">1.05</span><span class="hljs-punctuation">,</span> x1 <span class="hljs-operator">=</span> <span class="hljs-number">1.35</span><span class="hljs-punctuation">,</span> y0 <span class="hljs-operator">=</span> <span class="hljs-number">1.06</span><span class="hljs-punctuation">,</span> y1<span class="hljs-operator">=</span><span class="hljs-number">1</span><span class="hljs-punctuation">,</span> text <span class="hljs-operator">=</span> <span class="hljs-string">'type 4 cells'</span><span class="hljs-punctuation">,</span><br> rectArgs <span class="hljs-operator">=</span> <span class="hljs-built_in">list</span><span class="hljs-punctuation">(</span>border<span class="hljs-operator">=</span><span class="hljs-string">'#F9F2E4'</span><span class="hljs-punctuation">,</span>lwd<span class="hljs-operator">=</span><span class="hljs-number">0.5</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><br> textArgs <span class="hljs-operator">=</span> <span class="hljs-built_in">list</span><span class="hljs-punctuation">(</span>col<span class="hljs-operator">=</span><span class="hljs-string">'black'</span><span class="hljs-punctuation">,</span>cex <span class="hljs-operator">=</span> <span class="hljs-number">1</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br><br><br><span class="hljs-comment">#plot group#</span><br>Idents<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">)</span> <span class="hljs-operator"><-</span> <span class="hljs-string">"SCT_snn_res.0.1"</span><br>col_use <span class="hljs-operator"><-</span> my36colors<span class="hljs-punctuation">[</span><span class="hljs-number">1</span><span class="hljs-operator">:</span>nlevels<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">)</span><span class="hljs-punctuation">]</span><br>cc <span class="hljs-operator"><-</span> get_metadata<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">,</span> color <span class="hljs-operator">=</span> col_use<span class="hljs-punctuation">)</span><br>cc <span class="hljs-operator">%>%</span><br> dplyr<span class="hljs-operator">::</span>group_by<span class="hljs-punctuation">(</span><span class="hljs-string">"SCT_snn_res.0.1"</span><span class="hljs-punctuation">)</span> <span class="hljs-operator">%>%</span><br> summarize<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> median<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> x<span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span>y <span class="hljs-operator">=</span> median<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> y<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span> <span class="hljs-operator">-></span> centers<br>col_group <span class="hljs-operator"><-</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-string">'#bff542'</span><span class="hljs-punctuation">,</span><span class="hljs-string">'#83f78f'</span><span class="hljs-punctuation">,</span><span class="hljs-string">'#EBA1A2'</span><span class="hljs-punctuation">,</span><span class="hljs-string">'#D70016'</span><span class="hljs-punctuation">)</span><br>lgd_points <span class="hljs-operator">=</span> Legend<span class="hljs-punctuation">(</span>labels <span class="hljs-operator">=</span> <span class="hljs-built_in">names</span><span class="hljs-punctuation">(</span>table<span class="hljs-punctuation">(</span>cc<span class="hljs-operator">$</span>`SCT_snn_res.0.1`<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> type <span class="hljs-operator">=</span> <span class="hljs-string">"points"</span><span class="hljs-punctuation">,</span> <br> title_position <span class="hljs-operator">=</span> <span class="hljs-string">"topleft"</span><span class="hljs-punctuation">,</span> <br> title <span class="hljs-operator">=</span> <span class="hljs-string">"Group"</span><span class="hljs-punctuation">,</span><br> title_gp <span class="hljs-operator">=</span> gpar<span class="hljs-punctuation">(</span>col<span class="hljs-operator">=</span><span class="hljs-string">'black'</span><span class="hljs-punctuation">,</span>fontsize <span class="hljs-operator">=</span> <span class="hljs-number">7</span><span class="hljs-punctuation">,</span> fontface<span class="hljs-operator">=</span><span class="hljs-string">'bold'</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><br> legend_gp <span class="hljs-operator">=</span> gpar<span class="hljs-punctuation">(</span>col <span class="hljs-operator">=</span> col_group<span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><br> labels_gp <span class="hljs-operator">=</span> gpar<span class="hljs-punctuation">(</span>col<span class="hljs-operator">=</span><span class="hljs-string">'black'</span><span class="hljs-punctuation">,</span>fontsize <span class="hljs-operator">=</span> <span class="hljs-number">5</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><br> grid_height <span class="hljs-operator">=</span> unit<span class="hljs-punctuation">(</span><span class="hljs-number">2</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"mm"</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><br> grid_width <span class="hljs-operator">=</span> unit<span class="hljs-punctuation">(</span><span class="hljs-number">2</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"mm"</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><br> background <span class="hljs-operator">=</span> col_group<span class="hljs-punctuation">)</span><br>draw<span class="hljs-punctuation">(</span>lgd_points<span class="hljs-punctuation">,</span> x <span class="hljs-operator">=</span> unit<span class="hljs-punctuation">(</span><span class="hljs-number">15</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"mm"</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> y <span class="hljs-operator">=</span> unit<span class="hljs-punctuation">(</span><span class="hljs-number">50</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"mm"</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><br> just <span class="hljs-operator">=</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-string">"right"</span><span class="hljs-punctuation">,</span> <span class="hljs-string">"bottom"</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br><br>dev.off<span class="hljs-punctuation">(</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">}</span> <span class="hljs-comment"># 导出 PDF 结束</span><br></code></pre></td></tr></table></figure>
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