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   "cell_type": "markdown",
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   "source": [
    "## Overview\n",
    "\n",
    "This notebook demonstrates how to use a Monet model to perform a label transfer between datasets. This requires one \"reference\" dataset for which labels are already available (usually from a clustering analysis), and a second \"target\" dataset to which the labels are supposed to be transfered. Usually the reference and target datasets originate from the same tissue type.\n",
    "\n",
    "Very briefly, the way the label transfer works is by first projecting both datasets into a shared PC space, defined by a Monet model. Then, a K-nearest-neighbor classifier is trained on the reference data (cells + labels). Finally, the trained classifier is used to predict the cell types of the cells in teh target dataset. For more details, see the [Monet paper (Wagner, 2020)](https://www.biorxiv.org/content/10.1101/2020.06.08.140673v2).\n",
    "\n",
    "For this tutorial, we will use the same two PBMC datasets as in the previous tutorial. These are two datasets generated using two different technologies/chemistries (10x Genomics' Chromium v2 and v3). This example is also shown and discussed in more detail in Figure 4 of the [Monet paper (Wagner, 2020)](https://www.biorxiv.org/content/10.1101/2020.06.08.140673v2)."
   ]
  },
  {
   "cell_type": "markdown",
   "metadata": {},
   "source": [
    "### Set up the notebook"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 1,
   "metadata": {},
   "outputs": [
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      "text/html": [
       "<style>\n",
       "    /* source: http://stackoverflow.com/a/24207353 */\n",
       "    .container { width:95% !important; }\n",
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       "    </style>"
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       "        <script type=\"text/javascript\">\n",
       "        window.PlotlyConfig = {MathJaxConfig: 'local'};\n",
       "        if (window.MathJax) {MathJax.Hub.Config({SVG: {font: \"STIX-Web\"}});}\n",
       "        if (typeof require !== 'undefined') {\n",
       "        require.undef(\"plotly\");\n",
       "        requirejs.config({\n",
       "            paths: {\n",
       "                'plotly': ['https://cdn.plot.ly/plotly-latest.min']\n",
       "            }\n",
       "        });\n",
       "        require(['plotly'], function(Plotly) {\n",
       "            window._Plotly = Plotly;\n",
       "        });\n",
       "        }\n",
       "        </script>\n",
       "        "
      ]
     },
     "metadata": {},
     "output_type": "display_data"
    }
   ],
   "source": [
    "# change notebook width and font\n",
    "from IPython.core.display import HTML, display\n",
    "display(HTML(\"\"\"<style>\n",
    "    /* source: http://stackoverflow.com/a/24207353 */\n",
    "    .container { width:95% !important; }\n",
    "    div.prompt, div.CodeMirror pre, div.output_area pre { font-family:'Hack', monospace; font-size: 10.5pt; }\n",
    "    </style>\"\"\"))\n",
    "\n",
    "from monet import util\n",
    "\n",
    "_LOGGER = util.configure_logger()\n",
    "\n",
    "# the following is to allow embedding of plotly figures\n",
    "from plotly.offline import init_notebook_mode\n",
    "import plotly.graph_objs as go\n",
    "init_notebook_mode(connected=True)"
   ]
  },
  {
   "cell_type": "markdown",
   "metadata": {},
   "source": [
    "## Perform label transfer\n",
    "\n",
    "The label transfer method is provided by Monet in the `label_transfer.transfer_knn()` function. By default, the K-nearest neighbor classifier uses *K=20*. You can change this by passing the `num_neighbors` parameter to the `transfer_knn()` function."
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 2,
   "metadata": {},
   "outputs": [
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "[2020-06-17 13:02:32] (monet.util.files) INFO: Loaded labels for 10681 cells from tab-delimited plain-text file.\n",
      "Monocytes                     3345\n",
      "CD4+ Memory T cells           1847\n",
      "Naive T cells                 1361\n",
      "Naive B cells                  956\n",
      "Other                          888\n",
      "CD8+/CD161+ Memory T cells     669\n",
      "NK cells                       563\n",
      "Memory B cells                 457\n",
      "CD8+/CD161- Memory T cells     363\n",
      "mDCs                           151\n",
      "pDCs                            81\n",
      "Name: 0, dtype: int64\n",
      "[2020-06-17 13:02:32] (monet.latent.monet_model) INFO: Loaded Monet model from pickle file \"data/v3_human_pbmc_10k_monet_model.pickle\".\n",
      "[2020-06-17 13:02:35] (monet.core.exp_matrix) INFO: Loaded expression matrix with 10681 cells and 16319 genes -- .npz format, 36.7 MB (hash: f9d7fac20f4de6184ff55388c267699a).\n",
      "[2020-06-17 13:02:38] (monet.core.exp_matrix) INFO: Loaded expression matrix with 8381 cells and 15510 genes -- .npz format, 19.9 MB (hash: c299645ab748c9dbe4030fc4cace369b).\n",
      "[2020-06-17 13:02:39] (monet.latent.pca_model) INFO: Expression profiles will be scaled 1.00x (on average).\n",
      "[2020-06-17 13:02:44] (monet.latent.pca_model) INFO: Projection onto 30 PCs retained 32.1 % of the total variance in the scaled and FT-transformed data.\n",
      "[2020-06-17 13:02:44] (monet.latent.pca_model) WARNING: No expression data for 1153 / 15510 genes (7.4 %) in the PCA model.\n",
      "[2020-06-17 13:02:45] (monet.latent.pca_model) INFO: Expression profiles will be scaled 1.57x (on average).\n",
      "[2020-06-17 13:02:49] (monet.latent.pca_model) INFO: Projection onto 30 PCs retained 20.8 % of the total variance in the scaled and FT-transformed data.\n",
      "Naive T cells                 2116\n",
      "Monocytes                     1670\n",
      "CD4+ Memory T cells           1405\n",
      "CD8+/CD161- Memory T cells     791\n",
      "Naive B cells                  781\n",
      "Other                          557\n",
      "Memory B cells                 370\n",
      "NK cells                       280\n",
      "mDCs                           188\n",
      "CD8+/CD161+ Memory T cells     161\n",
      "pDCs                            62\n",
      "dtype: int64\n"
     ]
    },
    {
     "data": {
      "text/plain": [
       "24"
      ]
     },
     "execution_count": 2,
     "metadata": {},
     "output_type": "execute_result"
    }
   ],
   "source": [
    "import gc\n",
    "\n",
    "from monet import ExpMatrix\n",
    "from monet import MonetModel\n",
    "from monet import label_transfer\n",
    "from monet import util\n",
    "\n",
    "import pandas as pd\n",
    "\n",
    "monet_model_file = 'data/v3_human_pbmc_10k_monet_model.pickle'\n",
    "ref_expression_file = 'data/v3_human_pbmc_10k_expression.npz'\n",
    "target_expression_file = 'data/v2_human_pbmc_8k_expression.npz'\n",
    "\n",
    "ref_cell_label_file = 'data/v3_human_pbmc_10k_clustering_annotated.tsv'\n",
    "\n",
    "ref_cell_labels = util.load_cell_labels(ref_cell_label_file)\n",
    "print(ref_cell_labels.value_counts())\n",
    "\n",
    "monet_model = MonetModel.load_pickle(monet_model_file)\n",
    "\n",
    "ref_matrix = ExpMatrix.load_npz(ref_expression_file)\n",
    "target_matrix = ExpMatrix.load_npz(target_expression_file)\n",
    "\n",
    "target_cell_labels = label_transfer.transfer_knn(\n",
    "    monet_model, ref_matrix, ref_cell_labels, target_matrix)\n",
    "print(target_cell_labels.value_counts())\n",
    "\n",
    "# free up memory\n",
    "del ref_matrix, target_matrix; gc.collect()"
   ]
  },
  {
   "cell_type": "markdown",
   "metadata": {},
   "source": [
    "## Plot the results"
   ]
  },
  {
   "cell_type": "code",
   "execution_count": 3,
   "metadata": {},
   "outputs": [
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "[2020-06-17 13:02:53] (monet.core.exp_matrix) INFO: Loaded expression matrix with 8381 cells and 15510 genes -- .npz format, 19.9 MB (hash: c299645ab748c9dbe4030fc4cace369b).\n",
      "[2020-06-17 13:02:53] (root) INFO: No Monet model provided, performing PCA to determine first 30principal components...\n",
      "[2020-06-17 13:02:53] (monet.latent.pca_model) INFO: Converted matrix to float32 data type.\n",
      "[2020-06-17 13:02:58] (monet.latent.pca_model) INFO: The PCA took 1.4 s.\n",
      "[2020-06-17 13:02:58] (monet.latent.pca_model) INFO: The fraction of variance explained by the 30 selected PCs is 25.5 %.\n",
      "[2020-06-17 13:02:58] (root) INFO: Performing t-SNE...\n",
      "[2020-06-17 13:03:18] (root) INFO: t-SNE took 20.2 s.\n"
     ]
    },
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       "            console.log([gd, 'removed!']);\n",
       "            Plotly.purge(gd);\n",
       "            observer.disconnect();\n",
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       "\n",
       "// Listen for the removal of the full notebook cells\n",
       "var notebookContainer = gd.closest('#notebook-container');\n",
       "if (notebookContainer) {{\n",
       "    x.observe(notebookContainer, {childList: true});\n",
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       "\n",
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       "var outputEl = gd.closest('.output');\n",
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       "            </script>\n",
       "        </div>"
      ]
     },
     "metadata": {},
     "output_type": "display_data"
    },
    {
     "name": "stdout",
     "output_type": "stream",
     "text": [
      "Naive T cells                 2116\n",
      "Monocytes                     1670\n",
      "CD4+ Memory T cells           1405\n",
      "CD8+/CD161- Memory T cells     791\n",
      "Naive B cells                  781\n",
      "Other                          557\n",
      "Memory B cells                 370\n",
      "NK cells                       280\n",
      "mDCs                           188\n",
      "CD8+/CD161+ Memory T cells     161\n",
      "pDCs                            62\n",
      "dtype: int64\n"
     ]
    },
    {
     "data": {
      "text/plain": [
       "8251"
      ]
     },
     "execution_count": 3,
     "metadata": {},
     "output_type": "execute_result"
    }
   ],
   "source": [
    "import gc\n",
    "\n",
    "from monet import ExpMatrix\n",
    "from monet import visualize\n",
    "\n",
    "expression_file = 'data/v2_human_pbmc_8k_expression.npz'\n",
    "\n",
    "matrix = ExpMatrix.load_npz(expression_file)\n",
    "\n",
    "cluster_order = [\n",
    "    'Naive T cells',\n",
    "    'CD4+ Memory T cells',\n",
    "    'CD8+/CD161- Memory T cells',\n",
    "    'CD8+/CD161+ Memory T cells',\n",
    "    'NK cells',\n",
    "    'Naive B cells',\n",
    "    'Memory B cells',\n",
    "    'Monocytes',\n",
    "    'mDCs',\n",
    "    'pDCs',\n",
    "    'Other',\n",
    "]\n",
    "    \n",
    "cluster_colors = {\n",
    "    'Other': 'lightgray',\n",
    "}\n",
    "\n",
    "fig, tsne_scores = visualize.tsne_plot(\n",
    "    matrix, num_components=30,\n",
    "    cell_labels=target_cell_labels,\n",
    "    cluster_order=cluster_order,\n",
    "    cluster_colors=cluster_colors,\n",
    "    width=1200)\n",
    "fig.show()\n",
    "\n",
    "print(target_cell_labels.value_counts())\n",
    "\n",
    "# free up memory\n",
    "del matrix; gc.collect()"
   ]
  }
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