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- # Dash app to visualize scRNA-seq data quality control metrics from scanpy objects
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- # Shoutout to Coding-with-Adam for the initial template of the project:
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- # https://github.com/Coding-with-Adam/Dash-by-Plotly/blob/master/Dash%20Components/Graph/dash-graph.py
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-
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- import dash
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- from dash import dcc, html, Output, Input, callback
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- import plotly.express as px
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- import dash_callback_chain
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- import yaml
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- import polars as pl
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- import os
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- pl.enable_string_cache(False)
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-
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- dash.register_page(__name__, location="sidebar")
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-
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- dataset = "data10xflex/corg/10xflexcorg_umap_clusres"
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-
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- # Set custom resolution for plots:
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- config_fig = {
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- 'toImageButtonOptions': {
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- 'format': 'svg',
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- 'filename': 'custom_image',
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- 'height': 600,
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- 'width': 700,
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- 'scale': 1,
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- }
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- }
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- from adlfs import AzureBlobFileSystem
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- mountpount=os.environ['AZURE_MOUNT_POINT'],
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- AZURE_STORAGE_ACCESS_KEY=os.getenv('AZURE_STORAGE_ACCESS_KEY')
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- AZURE_STORAGE_ACCOUNT=os.getenv('AZURE_STORAGE_ACCOUNT')
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-
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- # Load in config file
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- config_path = "./data/config.yaml"
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-
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- # Add the read-in data from the yaml file
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- def read_config(filename):
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- with open(filename, 'r') as yaml_file:
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- config = yaml.safe_load(yaml_file)
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- return config
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-
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- config = read_config(config_path)
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- path_parquet = config.get("path_parquet")
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- col_batch = config.get("col_batch")
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- col_features = config.get("col_features")
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- col_counts = config.get("col_counts")
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- col_mt = config.get("col_mt")
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-
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- #filepath = f"az://{path_parquet}"
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-
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- storage_options={'account_name': AZURE_STORAGE_ACCOUNT, 'account_key': AZURE_STORAGE_ACCESS_KEY,'anon': False}
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- #azfs = AzureBlobFileSystem(**storage_options )
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-
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- # Load in multiple dataframes
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- df = pl.read_parquet(f"az://{dataset}.parquet", storage_options=storage_options)
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-
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- # Setup the app
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- #external_stylesheets = ['https://codepen.io/chriddyp/pen/bWLwgP.css']
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- #app = dash.Dash(__name__, use_pages=True) #, requests_pathname_prefix='/dashboard1/'
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-
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- #df = pl.read_parquet(filepath,storage_options=storage_options)
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- #df = pl.DataFrame()
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- #abfs = AzureBlobFileSystem(account_name=accountname,account_key=accountkey)
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- #df = df.rename({"__index_level_0__": "Unnamed: 0"})
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-
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- #df1 = pl.read_parquet(filepath, storage_options=storage_options)
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-
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- #df2 = pl.read_parquet(f"az://data10xflex/{dataset_chosen}.parquet", storage_options=storage_options)
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-
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- #tab0_content = html.Div([
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- # html.Label("Dataset chosen"),
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- # dcc.Dropdown(id='dpdn1', value="corg/10xflexcorg_umap_clusres", multi=False,
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- # options=["corg/10xflexcorg_umap_clusres","d1011/10xflexd1011_umap_clusres"])
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- #])
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-
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- #@app.callback(
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- # Input(component_id='dpdn1', component_property='value')
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- #)
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-
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- #def update_filepath(dpdn1):
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- # global df
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- # if str(f"az://data10xflex/{dpdn1}.parquet") != str(filepath):
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- # print("not identical filepath, chosing other")
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- # df2 = pl.read_parquet(f"az://data10xflex/{dpdn1}.parquet", storage_options=storage_options)
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- # df = df2
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- # return
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-
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- #df = pl.read_parquet(filepath, storage_options=storage_options)
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- min_value = df[col_features].min()
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- max_value = df[col_features].max()
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-
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- min_value_2 = df[col_counts].min()
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- min_value_2 = round(min_value_2)
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- max_value_2 = df[col_counts].max()
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- max_value_2 = round(max_value_2)
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-
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- min_value_3 = df[col_mt].min()
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- min_value_3 = round(min_value_3, 1)
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- max_value_3 = df[col_mt].max()
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- max_value_3 = round(max_value_3, 1)
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-
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- # Loads in the conditions specified in the yaml file
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-
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- # Note: Future version perhaps all values from a column in the dataframe of the parquet file
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- # Note 2: This could also be a tsv of the categories and own specified colors
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- #conditions = df[col_batch].unique().to_list()
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- # Create the first tab content
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- # Add Sliders for three QC params: N genes by counts, total amount of reads and pct MT reads
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-
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- tab1_content = html.Div([
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- html.Label("Column chosen"),
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- dcc.Dropdown(id='dpdn2', value="batch", multi=False,
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- options=df.columns),
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- html.Label("N Genes by Counts"),
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- dcc.RangeSlider(
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- id='range-slider-1',
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- step=250,
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- value=[min_value, max_value],
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- marks={i: str(i) for i in range(min_value, max_value + 1, 250)},
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- ),
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- dcc.Input(id='min-slider-1', type='number', value=min_value, debounce=True),
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- dcc.Input(id='max-slider-1', type='number', value=max_value, debounce=True),
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- html.Label("Total Counts"),
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- dcc.RangeSlider(
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- id='range-slider-2',
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- step=7500,
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- value=[min_value_2, max_value_2],
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- marks={i: str(i) for i in range(min_value_2, max_value_2 + 1, 7500)},
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- ),
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- dcc.Input(id='min-slider-2', type='number', value=min_value_2, debounce=True),
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- dcc.Input(id='max-slider-2', type='number', value=max_value_2, debounce=True),
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- html.Label("Percent Mitochondrial Genes"),
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- dcc.RangeSlider(
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- id='range-slider-3',
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- step=5,
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- min=0,
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- max=100,
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- value=[min_value_3, max_value_3],
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- ),
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- dcc.Input(id='min-slider-3', type='number', value=min_value_3, debounce=True),
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- dcc.Input(id='max-slider-3', type='number', value=max_value_3, debounce=True),
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- html.Div([
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- dcc.Graph(id='pie-graph', figure={}, className='four columns',config=config_fig),
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- dcc.Graph(id='my-graph', figure={}, clickData=None, hoverData=None,
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- className='four columns',config=config_fig
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- ),
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- dcc.Graph(id='scatter-plot', figure={}, className='four columns',config=config_fig)
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- ]),
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- html.Div([
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- dcc.Graph(id='scatter-plot-2', figure={}, className='four columns',config=config_fig)
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- ]),
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- html.Div([
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- dcc.Graph(id='scatter-plot-3', figure={}, className='four columns',config=config_fig)
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- ]),
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- html.Div([
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- dcc.Graph(id='scatter-plot-4', figure={}, className='four columns',config=config_fig)
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- ]),
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- ])
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-
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- # Create the second tab content with scatter-plot-5 and scatter-plot-6
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- tab2_content = html.Div([
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- html.Div([
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- html.Label("S-cycle genes"),
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- dcc.Dropdown(id='dpdn3', value="MCM5", multi=False,
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- options=[
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- "MCM5",
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- "PCNA",
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- "TYMS",
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- "FEN1",
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- "MCM2",
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- "MCM4",
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- "RRM1",
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- "UNG",
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- "GINS2",
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- "MCM6",
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- "CDCA7",
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- "DTL",
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- "PRIM1",
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- "UHRF1",
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- "MLF1IP",
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- "HELLS",
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- "RFC2",
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- "RPA2",
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- "NASP",
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- "RAD51AP1",
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- "GMNN",
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- "WDR76",
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- "SLBP",
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- "CCNE2",
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- "UBR7",
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- "POLD3",
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- "MSH2",
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- "ATAD2",
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- "RAD51",
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- "RRM2",
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- "CDC45",
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- "CDC6",
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- "EXO1",
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- "TIPIN",
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- "DSCC1",
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- "BLM",
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- "CASP8AP2",
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- "USP1",
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- "CLSPN",
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- "POLA1",
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- "CHAF1B",
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- "BRIP1",
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- "E2F8"
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- ]),
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- html.Label("G2M-cycle genes"),
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- dcc.Dropdown(id='dpdn4', value="TOP2A", multi=False,
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- options=[
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- 'HMGB2', 'CDK1', 'NUSAP1', 'UBE2C', 'BIRC5', 'TPX2', 'TOP2A', 'NDC80', 'CKS2', 'NUF2', 'CKS1B', 'MKI67', 'TMPO', 'CENPF', 'TACC3', 'FAM64A', 'SMC4', 'CCNB2', 'CKAP2L', 'CKAP2', 'AURKB', 'BUB1', 'KIF11', 'ANP32E', 'TUBB4B', 'GTSE1', 'KIF20B', 'HJURP', 'CDCA3', 'HN1', 'CDC20', 'TTK', 'CDC25C', 'KIF2C', 'RANGAP1', 'NCAPD2', 'DLGAP5', 'CDCA2', 'CDCA8', 'ECT2', 'KIF23', 'HMMR', 'AURKA', 'PSRC1', 'ANLN', 'LBR', 'CKAP5',
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- 'CENPE', 'CTCF', 'NEK2', 'G2E3', 'GAS2L3', 'CBX5', 'CENPA'
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- ]),
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- ]),
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- html.Div([
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- dcc.Graph(id='scatter-plot-5', figure={}, className='three columns',config=config_fig)
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- ]),
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- html.Div([
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- dcc.Graph(id='scatter-plot-6', figure={}, className='three columns',config=config_fig)
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- ]),
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- html.Div([
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- dcc.Graph(id='scatter-plot-7', figure={}, className='three columns',config=config_fig)
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- ]),
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- html.Div([
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- dcc.Graph(id='scatter-plot-8', figure={}, className='three columns',config=config_fig)
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- ]),
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- ])
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-
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- # Create the second tab content with scatter-plot-5 and scatter-plot-6
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- tab3_content = html.Div([
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- html.Div([
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- html.Label("UMAP condition 1"),
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- dcc.Dropdown(id='dpdn5', value="batch", multi=False,
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- options=df.columns),
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- html.Label("UMAP condition 2"),
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- dcc.Dropdown(id='dpdn6', value="n_genes_by_counts", multi=False,
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- options=df.columns),
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- html.Div([
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- dcc.Graph(id='scatter-plot-9', figure={}, className='four columns',config=config_fig)
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- ]),
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- html.Div([
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- dcc.Graph(id='scatter-plot-10', figure={}, className='four columns',config=config_fig)
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- ]),
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- html.Div([
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- dcc.Graph(id='scatter-plot-11', figure={}, className='four columns',config=config_fig)
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- ]),
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- html.Div([
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- dcc.Graph(id='my-graph2', figure={}, clickData=None, hoverData=None,
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- className='four columns',config=config_fig
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- )
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- ]),
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- ]),
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- ])
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- # html.Div([
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- # dcc.Graph(id='scatter-plot-12', figure={}, className='four columns',config=config_fig)
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- # ]),
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-
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-
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- tab4_content = html.Div([
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- html.Div([
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- html.Label("Multi gene"),
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- dcc.Dropdown(id='dpdn7', value=["PAX6","TP63","S100A9"], multi=True,
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- options=df.columns),
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- ]),
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- html.Div([
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- dcc.Graph(id='scatter-plot-12', figure={}, className='four columns',config=config_fig)
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- ]),
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- ])
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-
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- # Define the tabs layout
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- layout = html.Div([
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- dcc.Tabs(id='tabs', style= {'width': 600,
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- 'font-size': '100%',
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- 'height': 50}, value='tab1',children=[
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- #dcc.Tab(label='Dataset', value='tab0', children=tab0_content),
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- dcc.Tab(label='QC', value='tab1', children=tab1_content),
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- dcc.Tab(label='Cell cycle', value='tab2', children=tab2_content),
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- dcc.Tab(label='Custom', value='tab3', children=tab3_content),
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- dcc.Tab(label='Multi dot', value='tab4', children=tab4_content),
282
- ]),
283
- ])
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-
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- # Define the circular callback
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- @callback(
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- Output("min-slider-1", "value"),
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- Output("max-slider-1", "value"),
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- Output("min-slider-2", "value"),
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- Output("max-slider-2", "value"),
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- Output("min-slider-3", "value"),
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- Output("max-slider-3", "value"),
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- Input("min-slider-1", "value"),
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- Input("max-slider-1", "value"),
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- Input("min-slider-2", "value"),
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- Input("max-slider-2", "value"),
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- Input("min-slider-3", "value"),
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- Input("max-slider-3", "value"),
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- )
300
- def circular_callback(min_1, max_1, min_2, max_2, min_3, max_3):
301
- return min_1, max_1, min_2, max_2, min_3, max_3
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-
303
- @callback(
304
- Output('range-slider-1', 'value'),
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- Output('range-slider-2', 'value'),
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- Output('range-slider-3', 'value'),
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- Input('min-slider-1', 'value'),
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- Input('max-slider-1', 'value'),
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- Input('min-slider-2', 'value'),
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- Input('max-slider-2', 'value'),
311
- Input('min-slider-3', 'value'),
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- Input('max-slider-3', 'value'),
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- )
314
- def update_slider_values(min_1, max_1, min_2, max_2, min_3, max_3):
315
- return [min_1, max_1], [min_2, max_2], [min_3, max_3]
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-
317
- @callback(
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- Output(component_id='my-graph', component_property='figure'),
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- Output(component_id='pie-graph', component_property='figure'),
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- Output(component_id='scatter-plot', component_property='figure'),
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- Output(component_id='scatter-plot-2', component_property='figure'),
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- Output(component_id='scatter-plot-3', component_property='figure'),
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- Output(component_id='scatter-plot-4', component_property='figure'), # Add this new scatter plot
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- Output(component_id='scatter-plot-5', component_property='figure'),
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- Output(component_id='scatter-plot-6', component_property='figure'),
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- Output(component_id='scatter-plot-7', component_property='figure'),
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- Output(component_id='scatter-plot-8', component_property='figure'),
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- Output(component_id='scatter-plot-9', component_property='figure'),
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- Output(component_id='scatter-plot-10', component_property='figure'),
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- Output(component_id='scatter-plot-11', component_property='figure'),
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- Output(component_id='scatter-plot-12', component_property='figure'),
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- Output(component_id='my-graph2', component_property='figure'),
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- Input(component_id='dpdn2', component_property='value'),
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- Input(component_id='dpdn3', component_property='value'),
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- Input(component_id='dpdn4', component_property='value'),
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- Input(component_id='dpdn5', component_property='value'),
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- Input(component_id='dpdn6', component_property='value'),
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- Input(component_id='dpdn7', component_property='value'),
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- Input(component_id='range-slider-1', component_property='value'),
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- Input(component_id='range-slider-2', component_property='value'),
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- Input(component_id='range-slider-3', component_property='value')
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- )
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-
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- def update_graph_and_pie_chart(col_chosen, s_chosen, g2m_chosen, condition1_chosen, condition2_chosen, condition3_chosen, range_value_1, range_value_2, range_value_3): #batch_chosen,
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- batch_chosen = df[col_chosen].unique().to_list()
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- dff = df.filter(
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- (pl.col(col_chosen).cast(str).is_in(batch_chosen)) &
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- (pl.col(col_features) >= range_value_1[0]) &
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- (pl.col(col_features) <= range_value_1[1]) &
350
- (pl.col(col_counts) >= range_value_2[0]) &
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- (pl.col(col_counts) <= range_value_2[1]) &
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- (pl.col(col_mt) >= range_value_3[0]) &
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- (pl.col(col_mt) <= range_value_3[1])
354
- )
355
-
356
- #Drop categories that are not in the filtered data
357
- dff = dff.with_columns(dff[col_chosen].cast(pl.Categorical))
358
-
359
- dff = dff.sort(col_chosen)
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-
361
- # Plot figures
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- fig_violin = px.violin(data_frame=dff, x=col_chosen, y=col_features, box=True, points="all",
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- color=col_chosen, hover_name=col_chosen,template="seaborn")
364
-
365
- # Cache commonly used subexpressions
366
- total_count = pl.lit(len(dff))
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- category_counts = dff.group_by(col_chosen).agg(pl.col(col_chosen).count().alias("count"))
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- category_counts = category_counts.with_columns(((pl.col("count") / total_count * 100).round(decimals=2)).alias("normalized_count"))
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-
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- # Sort the dataframe
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- #category_counts = category_counts.sort(col_chosen) does not work check if the names are different ...
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-
373
- # Display the result
374
- total_cells = total_count # Calculate total number of cells
375
- pie_title = f'Percentage of Total Cells: {total_cells}' # Include total cells in the title
376
-
377
- # Calculate the mean expression
378
-
379
- # Melt wide format DataFrame into long format
380
- # Specify batch column as string type and gene columns as float type
381
- list_conds = condition3_chosen
382
- list_conds += [col_chosen]
383
- dff_pre = dff.select(list_conds)
384
-
385
- # Melt wide format DataFrame into long format
386
- dff_long = dff_pre.melt(id_vars=col_chosen, variable_name="Gene", value_name="Mean expression")
387
-
388
- # Calculate the mean expression levels for each gene in each region
389
- expression_means = dff_long.lazy().group_by([col_chosen, "Gene"]).agg(pl.mean("Mean expression")).collect()
390
-
391
- # Calculate the percentage total expressed
392
- dff_long1 = dff_pre.melt(id_vars=col_chosen, variable_name="Gene")#.group_by(pl.all()).agg(pl.len())
393
- count = 1
394
- dff_long2 = dff_long1.with_columns(pl.lit(count).alias("len"))
395
- dff_long3 = dff_long2.filter(pl.col("value") > 0).group_by([col_chosen, "Gene"]).agg(pl.sum("len").alias("len"))
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- dff_long4 = dff_long2.group_by([col_chosen, "Gene"]).agg(pl.sum("len").alias("total"))
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- dff_5 = dff_long4.join(dff_long3, on=[col_chosen,"Gene"], how="outer")
398
- result = dff_5.select([
399
- pl.when((pl.col('len').is_not_null()) & (pl.col('total').is_not_null()))
400
- .then(pl.col('len') / pl.col('total')*100)
401
- .otherwise(None).alias("%"),
402
- ])
403
- result = result.with_columns(pl.col("%").fill_null(100))
404
- dff_5[["percentage"]] = result[["%"]]
405
- dff_5 = dff_5.select(pl.col(col_chosen,"Gene","percentage"))
406
-
407
- # Final part to join the percentage expressed and mean expression levels
408
- # TO DO
409
- expression_means = expression_means.join(dff_5, on=[col_chosen,"Gene"], how="inner")
410
-
411
- # Order the dataframe on ascending categories
412
- expression_means = expression_means.sort(col_chosen, descending=True)
413
-
414
- #expression_means = expression_means.select(["batch", "Gene", "Expression"] + condition3_chosen)
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- category_counts = category_counts.sort(col_chosen)
416
-
417
- fig_pie = px.pie(category_counts, values="normalized_count", names=col_chosen, labels=col_chosen, hole=.3, title=pie_title, template="seaborn")
418
-
419
- #labels = category_counts[col_chosen].to_list()
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- #values = category_counts["normalized_count"].to_list()
421
-
422
- # Create the scatter plots
423
- fig_scatter = px.scatter(data_frame=dff, x='X_umap-0', y='X_umap-1', color=col_chosen,
424
- labels={'X_umap-0': 'umap1' , 'X_umap-1': 'umap2'},
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- hover_name='batch',template="seaborn")
426
-
427
- fig_scatter_2 = px.scatter(data_frame=dff, x='X_umap-0', y='X_umap-1', color=col_mt,
428
- labels={'X_umap-0': 'umap1' , 'X_umap-1': 'umap2'},
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- hover_name='batch',template="seaborn")
430
-
431
- fig_scatter_3 = px.scatter(data_frame=dff, x='X_umap-0', y='X_umap-1', color=col_features,
432
- labels={'X_umap-0': 'umap1' , 'X_umap-1': 'umap2'},
433
- hover_name='batch',template="seaborn")
434
-
435
-
436
- fig_scatter_4 = px.scatter(data_frame=dff, x='X_umap-0', y='X_umap-1', color=col_counts,
437
- labels={'X_umap-0': 'umap1' , 'X_umap-1': 'umap2'},
438
- hover_name='batch',template="seaborn")
439
-
440
- fig_scatter_5 = px.scatter(data_frame=dff, x='X_umap-0', y='X_umap-1', color=s_chosen,
441
- labels={'X_umap-0': 'umap1' , 'X_umap-1': 'umap2'},
442
- hover_name='batch', title="S-cycle gene:",template="seaborn")
443
-
444
- fig_scatter_6 = px.scatter(data_frame=dff, x='X_umap-0', y='X_umap-1', color=g2m_chosen,
445
- labels={'X_umap-0': 'umap1' , 'X_umap-1': 'umap2'},
446
- hover_name='batch', title="G2M-cycle gene:",template="seaborn")
447
-
448
- fig_scatter_7 = px.scatter(data_frame=dff, x='X_umap-0', y='X_umap-1', color="S_score",
449
- labels={'X_umap-0': 'umap1' , 'X_umap-1': 'umap2'},
450
- hover_name='batch', title="S score:",template="seaborn")
451
-
452
- fig_scatter_8 = px.scatter(data_frame=dff, x='X_umap-0', y='X_umap-1', color="G2M_score",
453
- labels={'X_umap-0': 'umap1' , 'X_umap-1': 'umap2'},
454
- hover_name='batch', title="G2M score:",template="seaborn")
455
-
456
- # Sort values of custom in-between
457
- dff = dff.sort(condition1_chosen)
458
-
459
- fig_scatter_9 = px.scatter(data_frame=dff, x='X_umap-0', y='X_umap-1', color=condition1_chosen,
460
- labels={'X_umap-0': 'umap1' , 'X_umap-1': 'umap2'},
461
- hover_name='batch',template="seaborn")
462
-
463
- fig_scatter_10 = px.scatter(data_frame=dff, x='X_umap-0', y='X_umap-1', color=condition2_chosen,
464
- labels={'X_umap-0': 'umap1' , 'X_umap-1': 'umap2'},
465
- hover_name='batch',template="seaborn")
466
-
467
- fig_scatter_11 = px.scatter(data_frame=dff, x=condition1_chosen, y=condition2_chosen, color=condition1_chosen,
468
- #labels={'X_umap-0': 'umap1' , 'X_umap-1': 'umap2'},
469
- hover_name='batch',template="seaborn")
470
-
471
- fig_scatter_12 = px.scatter(data_frame=expression_means, x="Gene", y=col_chosen, color="Mean expression",
472
- size="percentage", size_max = 20,
473
- #labels={'X_umap-0': 'umap1' , 'X_umap-1': 'umap2'},
474
- hover_name=col_chosen,template="seaborn")
475
-
476
- fig_violin2 = px.violin(data_frame=dff, x=condition1_chosen, y=condition2_chosen, box=True, points="all",
477
- color=condition1_chosen, hover_name=condition1_chosen,template="seaborn")
478
-
479
-
480
- return fig_violin, fig_pie, fig_scatter, fig_scatter_2, fig_scatter_3, fig_scatter_4, fig_scatter_5, fig_scatter_6, fig_scatter_7, fig_scatter_8, fig_scatter_9, fig_scatter_10, fig_scatter_11, fig_scatter_12, fig_violin2
481
-
482
- # Set http://localhost:5000/ in web browser
483
- # Now create your regular FASTAPI application
484
-
485
- #if __name__ == '__main__':
486
- # app.run_server(debug=False, use_reloader=False, host='0.0.0.0', port=5000) #