{ "cells": [ { "cell_type": "markdown", "metadata": {}, "source": [ "# Plasma Treatment" ] }, { "cell_type": "code", "execution_count": 1, "metadata": { "collapsed": true }, "outputs": [], "source": [ "from clustergrammer_widget import *\n", "net = Network(clustergrammer_widget)" ] }, { "cell_type": "code", "execution_count": 2, "metadata": { "collapsed": true }, "outputs": [], "source": [ "net.load_file('../cytof_data/Plasma_UCT.txt')\n", "plasma_df = net.export_df()" ] }, { "cell_type": "code", "execution_count": 4, "metadata": { "collapsed": true }, "outputs": [], "source": [ "# manually set treatment colors\n", "net.set_cat_color('col', 1, 'Marker-type: phospho marker', 'red')\n", "net.set_cat_color('col', 1, 'Marker-type: surface marker', 'blue')\n", "\n", "# manually set row colors: downsample\n", "net.set_cat_color('row', 2, 'Majority-Category: B cells', '#22316C')\n", "net.set_cat_color('row', 2, 'Majority-Category: Basophils', '#000033')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD14hi monocytes', 'yellow')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD14low monocytes', '#93b8bf')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD1c DCs', '#3636e2')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD4 Tcells', 'blue')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD4 Tcells_CD127hi', '#FF6347')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD4 Tcells CD161hi', '#F87531')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD4 Tcells_Tregs', '#8B4513')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD4 Tcells+CD27hi', '#330303')\n", "net.set_cat_color('row', 2, 'Majority-Category: CD8 Tcells', '#ffb247')\n", "net.set_cat_color('row', 2, 'Majority-Category: Neutrophils', 'purple')\n", "net.set_cat_color('row', 2, 'Majority-Category: NK cells_CD16hi', 'red')\n", "net.set_cat_color('row', 2, 'Majority-Category: NK cells_CD16hi_CD57hi', 'orange')\n", "net.set_cat_color('row', 2, 'Majority-Category: NK cells_CD56hi', '#e052e5')\n", "net.set_cat_color('row', 2, 'Majority-Category: Undefined', 'gray')\n", "\n", "# manually set row colors: subsample\n", "net.set_cat_color('row', 2, 'B cells', '#22316C')\n", "net.set_cat_color('row', 2, 'Basophils', '#000033')\n", "net.set_cat_color('row', 2, 'CD14hi monocytes', 'yellow')\n", "net.set_cat_color('row', 2, 'CD14low monocytes', '#93b8bf')\n", "net.set_cat_color('row', 2, 'CD1c DCs', '#3636e2')\n", "net.set_cat_color('row', 2, 'CD4 Tcells', 'blue')\n", "net.set_cat_color('row', 2, 'CD4 Tcells_CD127hi', '#FF6347')\n", "net.set_cat_color('row', 2, 'CD4 Tcells CD161hi', '#F87531')\n", "net.set_cat_color('row', 2, 'CD4 Tcells_Tregs', '#8B4513')\n", "net.set_cat_color('row', 2, 'CD4 Tcells+CD27hi', '#330303')\n", "net.set_cat_color('row', 2, 'CD8 Tcells', '#ffb247')\n", "net.set_cat_color('row', 2, 'Neutrophils', 'purple')\n", "net.set_cat_color('row', 2, 'NK cells_CD16hi', 'red')\n", "net.set_cat_color('row', 2, 'NK cells_CD16hi_CD57hi', 'orange')\n", "net.set_cat_color('row', 2, 'NK cells_CD56hi', '#e052e5')\n", "net.set_cat_color('row', 2, 'Undefined', 'gray')\n", "\n", "# manually set treatment colors\n", "net.set_cat_color('row', 1, 'Majority-Treatment: Plasma', 'blue')\n", "net.set_cat_color('row', 1, 'Majority-Treatment: PMA', 'red')\n", "\n", "net.set_cat_color('row', 1, 'Treatment: Plasma', 'blue')\n", "net.set_cat_color('row', 1, 'Treatment: PMA', 'red')" ] }, { "cell_type": "markdown", "metadata": { "collapsed": true }, "source": [ "# Plasma Subsample All Markers" ] }, { "cell_type": "code", "execution_count": 6, "metadata": {}, "outputs": [ { "data": { "application/vnd.jupyter.widget-view+json": { "model_id": "c83cd419105f4a1f8c9fc2a9f85fea99" } }, "metadata": {}, "output_type": "display_data" } ], "source": [ "net.load_df(plasma_df)\n", "net.normalize(axis='col', norm_type='zscore', keep_orig=False)\n", "\n", "# subsample the data so that both treatments have the same number of cells\n", "net.random_sample(axis='row', num_samples=1000, random_state=99)\n", "\n", "# clip z-scores since we do not care about extreme outliers\n", "net.clip(-10,10)\n", "\n", "net.cluster(views=[])\n", "net.widget()" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "# Plasma Downsample All Markers" ] }, { "cell_type": "code", "execution_count": 7, "metadata": {}, "outputs": [ { "data": { "application/vnd.jupyter.widget-view+json": { "model_id": "e13f2a28eb094a0d82aac4ad66f8ab29" } }, "metadata": {}, "output_type": "display_data" } ], "source": [ "net.load_df(plasma_df)\n", "net.normalize(axis='col', norm_type='zscore', keep_orig=False)\n", "# downsample to 1000 clusters\n", "ds_data = net.downsample(ds_type='kmeans', axis='row', num_samples=1000)\n", "# clip z-scores since we do not care about extreme outliers\n", "net.clip(-10,10)\n", "net.cluster(views=[])\n", "net.widget()" ] }, { "cell_type": "markdown", "metadata": { "collapsed": true }, "source": [ "# Plasma Phopho Markers Subsample" ] }, { "cell_type": "code", "execution_count": 8, "metadata": {}, "outputs": [ { "data": { "application/vnd.jupyter.widget-view+json": { "model_id": "9bce63d31ea04e4795206386372584a8" } }, "metadata": {}, "output_type": "display_data" } ], "source": [ "net.load_df(plasma_df)\n", "net.filter_cat('col', 1, 'Marker-type: phospho marker')\n", "net.normalize(axis='col', norm_type='zscore', keep_orig=False)\n", "net.random_sample(axis='row', num_samples=1000, random_state=1000)\n", "net.clip(-10,10)\n", "net.cluster(views=[])\n", "net.widget()" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "# Plasma Phospho Markers Downsample" ] }, { "cell_type": "code", "execution_count": 9, "metadata": {}, "outputs": [ { "data": { "application/vnd.jupyter.widget-view+json": { "model_id": "b67db7f00b10475c889281c24eca81af" } }, "metadata": {}, "output_type": "display_data" } ], "source": [ "net.load_df(plasma_df)\n", "net.filter_cat('col', 1, 'Marker-type: phospho marker')\n", "net.normalize(axis='col', norm_type='zscore', keep_orig=False)\n", "ds_data = net.downsample(ds_type='kmeans', axis='row', num_samples=1000)\n", "net.clip(-10,10)\n", "net.cluster(views=[])\n", "net.widget()" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "Similarly to our results clustering the combined Plasma and PMA treated cells, we again find a prominent cluster of CD14hi monocytes with high phosphorylation across several markers. However, the phosphorylation levels in this cluster are not as uniformly high as they are after PMA treatment." ] }, { "cell_type": "code", "execution_count": null, "metadata": { "collapsed": true }, "outputs": [], "source": [] } ], "metadata": { "anaconda-cloud": {}, "kernelspec": { "display_name": "Python [Root]", "language": "python", "name": "Python [Root]" }, "language_info": { "codemirror_mode": { "name": "ipython", "version": 2 }, "file_extension": ".py", "mimetype": "text/x-python", "name": "python", "nbconvert_exporter": "python", "pygments_lexer": "ipython2", "version": "2.7.12" }, "widgets": { "application/vnd.jupyter.widget-state+json": { "state": { "74abb70e403f468daa42b65f916aa9a4": { "model_module": "jupyter-js-widgets", "model_module_version": "~2.1.4", "model_name": "LayoutModel", "state": { "_model_module_version": "~2.1.4", "_view_module_version": "~2.1.4" } }, "7a89ea4344674d1792abf71ad9d3563f": { "model_module": "jupyter-js-widgets", "model_module_version": 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\"name\": \"Antibody: IkBa\", \"clust\": 0, \"cat_0_index\": 6, \"rank\": 2, \"cat-0\": \"Marker-type: phospho marker\", \"rankvar\": 3, \"ini\": 4}, {\"group\": [5.0, 5.0, 4.0, 4.0, 3.0, 2.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Antibody: pCREB\", \"clust\": 9, \"cat_0_index\": 7, \"rank\": 8, \"cat-0\": \"Marker-type: phospho marker\", \"rankvar\": 7, \"ini\": 3}, {\"group\": [7.0, 7.0, 6.0, 6.0, 5.0, 3.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Antibody: pERK1_2\", \"clust\": 6, \"cat_0_index\": 8, \"rank\": 3, \"cat-0\": \"Marker-type: phospho marker\", \"rankvar\": 4, \"ini\": 2}, {\"group\": [9.0, 9.0, 8.0, 8.0, 7.0, 5.0, 4.0, 3.0, 3.0, 1.0, 1.0], \"name\": \"Antibody: pS6\", \"clust\": 1, \"cat_0_index\": 9, \"rank\": 0, \"cat-0\": \"Marker-type: phospho marker\", \"rankvar\": 1, \"ini\": 1}]}", "viz_title": "updating python by restarting kernel" } }, "c83cd419105f4a1f8c9fc2a9f85fea99": { "model_module": "clustergrammer_widget", "model_module_version": "*", "model_name": 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\"Majority-Treatment: Plasma\", \"rankvar\": 638}, {\"group\": [77.0, 21.0, 7.0, 3.0, 1.0, 1.0, 1.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-934\", \"clust\": 78, \"cat_2_index\": 465, \"cat_1_index\": 879, \"cat_0_index\": 934, \"cat-2\": \"number in clust: 112\", \"rank\": 725, \"cat-1\": \"Majority-Category: NK cells_CD16hi\", \"ini\": 66, \"cat-0\": \"Majority-Treatment: Plasma\", \"rankvar\": 774}, {\"group\": [387.0, 105.0, 32.0, 14.0, 7.0, 2.0, 1.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-935\", \"clust\": 387, \"cat_2_index\": 29, \"cat_1_index\": 190, \"cat_0_index\": 935, \"cat-2\": \"number in clust: 14\", \"rank\": 691, \"cat-1\": \"Majority-Category: CD14hi monocytes\", \"ini\": 65, \"cat-0\": \"Majority-Treatment: Plasma\", \"rankvar\": 750}, {\"group\": [601.0, 179.0, 56.0, 21.0, 12.0, 5.0, 2.0, 1.0, 1.0, 1.0, 1.0], \"name\": \"Cluster: cluster-936\", \"clust\": 601, \"cat_2_index\": 985, \"cat_1_index\": 776, \"cat_0_index\": 936, \"cat-2\": \"number 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IkBa\", \"clust\": 3, \"cat_0_index\": 6, \"rank\": 2, \"cat-0\": \"Marker-type: phospho marker\", \"rankvar\": 4, \"ini\": 4}, {\"group\": [22.0, 21.0, 20.0, 18.0, 15.0, 13.0, 7.0, 2.0, 1.0, 1.0, 1.0], \"name\": \"Antibody: pCREB\", \"clust\": 27, \"cat_0_index\": 7, \"rank\": 27, \"cat-0\": \"Marker-type: phospho marker\", \"rankvar\": 11, \"ini\": 3}, {\"group\": [25.0, 24.0, 23.0, 20.0, 16.0, 13.0, 7.0, 2.0, 1.0, 1.0, 1.0], \"name\": \"Antibody: pERK1_2\", \"clust\": 23, \"cat_0_index\": 8, \"rank\": 15, \"cat-0\": \"Marker-type: phospho marker\", \"rankvar\": 3, \"ini\": 2}, {\"group\": [28.0, 27.0, 26.0, 23.0, 19.0, 16.0, 10.0, 2.0, 1.0, 1.0, 1.0], \"name\": \"Antibody: pS6\", \"clust\": 14, \"cat_0_index\": 9, \"rank\": 5, \"cat-0\": \"Marker-type: phospho marker\", \"rankvar\": 1, \"ini\": 1}]}", "viz_title": "updating python by restarting kernel" } } }, "version_major": 1, "version_minor": 0 } } }, "nbformat": 4, "nbformat_minor": 2 }