view SingleCellDataExtraction.py @ 3:c09e444635d9 draft

"planemo upload for repository https://github.com/ohsu-comp-bio/quantification commit 98cddbe59b6834a2368a0085b3938d5058f6d10b"

#Functions for reading in single cell imaging data
#Joshua Hess

#Import necessary modules
import skimage.io
import h5py
import pandas as pd
import numpy as np
import os
import skimage.measure as measure
import tifffile

from pathlib import Path

import sys


def gini_index(mask, intensity):
    x = intensity[mask]
    sorted_x = np.sort(x)
    n = len(x)
    cumx = np.cumsum(sorted_x, dtype=float)
    return (n + 1 - 2 * np.sum(cumx) / cumx[-1]) / n

def intensity_median(mask, intensity):
    return np.median(intensity[mask])

def MaskChannel(mask_loaded, image_loaded_z, intensity_props=["intensity_mean"]):
    """Function for quantifying a single channel image

    Returns a table with CellID according to the mask and the mean pixel intensity
    for the given channel for each cell"""
    # Look for regionprops in skimage
    builtin_props = set(intensity_props).intersection(measure._regionprops.PROP_VALS)
    # Otherwise look for them in this module
    extra_props = set(intensity_props).difference(measure._regionprops.PROP_VALS)
    dat = measure.regionprops_table(
        mask_loaded, image_loaded_z,
        properties = tuple(builtin_props),
        extra_properties = [globals()[n] for n in extra_props]
    )
    return dat


def MaskIDs(mask, mask_props=None):
    """This function will extract the CellIDs and the XY positions for each
    cell based on that cells centroid

    Returns a dictionary object"""

    all_mask_props = set(["label", "centroid", "area", "major_axis_length", "minor_axis_length", "eccentricity", "solidity", "extent", "orientation"])
    if mask_props is not None:
        all_mask_props = all_mask_props.union(mask_props)

    dat = measure.regionprops_table(
        mask,
        properties=all_mask_props
    )

    name_map = {
        "CellID": "label",
        "X_centroid": "centroid-1",
        "Y_centroid": "centroid-0",
        "Area": "area",
        "MajorAxisLength": "major_axis_length",
        "MinorAxisLength": "minor_axis_length",
        "Eccentricity": "eccentricity",
        "Solidity": "solidity",
        "Extent": "extent",
        "Orientation": "orientation",
    }
    for new_name, old_name in name_map.items():
        dat[new_name] = dat[old_name]
    for old_name in set(name_map.values()):
        del dat[old_name]

    return dat

def n_channels(image):
    """Returns the number of channel in the input image. Supports [OME]TIFF and HDF5."""

    image_path = Path(image)

    if image_path.suffix in ['.tiff', '.tif', '.btf']:
        s = tifffile.TiffFile(image).series[0]
        ndim = len(s.shape)
        if ndim == 2: return 1
        elif ndim == 3: return min(s.shape)
        else: raise Exception('mcquant supports only 2D/3D images.')

    elif image_path.suffix in ['.h5', '.hdf5']:
        f = h5py.File(image, 'r')
        dat_name = list(f.keys())[0]
        return f[dat_name].shape[3]

    else:
        raise Exception('mcquant currently supports [OME]TIFF and HDF5 formats only')

def PrepareData(image,z):
    """Function for preparing input for maskzstack function. Connecting function
    to use with mc micro ilastik pipeline"""

    image_path = Path(image)
    print(f'{image_path} at {z}', file=sys.stderr)

    #Check to see if image tif(f)
    if image_path.suffix in ['.tiff', '.tif', '.btf']:
        image_loaded_z = tifffile.imread(image, key=z)

    #Check to see if image is hdf5
    elif image_path.suffix in ['.h5', '.hdf5']:
        #Read the image
        f = h5py.File(image,'r')
        #Get the dataset name from the h5 file
        dat_name = list(f.keys())[0]
        #Retrieve the z^th channel
        image_loaded_z = f[dat_name][0,:,:,z]

    else:
        raise Exception('mcquant currently supports [OME]TIFF and HDF5 formats only')

    #Return the objects
    return image_loaded_z


def MaskZstack(masks_loaded,image,channel_names_loaded, mask_props=None, intensity_props=["intensity_mean"]):
    """This function will extract the stats for each cell mask through each channel
    in the input image

    mask_loaded: dictionary containing Tiff masks that represents the cells in your image.

    z_stack: Multichannel z stack image"""

    #Get the names of the keys for the masks dictionary
    mask_names = list(masks_loaded.keys())

    #Create empty dictionary to store channel results per mask
    dict_of_chan = {m_name: [] for m_name in mask_names}
    #Get the z channel and the associated channel name from list of channel names
    print(f'channels: {channel_names_loaded}', file=sys.stderr)
    print(f'num channels: {len(channel_names_loaded)}', file=sys.stderr)
    for z in range(len(channel_names_loaded)):
        #Run the data Prep function
        image_loaded_z = PrepareData(image,z)

        #Iterate through number of masks to extract single cell data
        for nm in range(len(mask_names)):
            #Use the above information to mask z stack
            dict_of_chan[mask_names[nm]].append(
                MaskChannel(masks_loaded[mask_names[nm]],image_loaded_z, intensity_props=intensity_props)
            )
        #Print progress
        print("Finished "+str(z))

    # Column order according to histoCAT convention (Move xy position to end with spatial information)
    last_cols = (
        "X_centroid",
        "Y_centroid",
        "column_centroid",
        "row_centroid",
        "Area",
        "MajorAxisLength",
        "MinorAxisLength",
        "Eccentricity",
        "Solidity",
        "Extent",
        "Orientation",
    )
    def col_sort(x):
        if x == "CellID":
            return -2
        try:
            return last_cols.index(x)
        except ValueError:
            return -1

    #Iterate through the masks and format quantifications for each mask and property
    for nm in mask_names:
        mask_dict = {}
        # Mean intensity is default property, stored without suffix
        mask_dict.update(
            zip(channel_names_loaded, [x["intensity_mean"] for x in dict_of_chan[nm]])
        )
        # All other properties are suffixed with their names
        for prop_n in set(dict_of_chan[nm][0].keys()).difference(["intensity_mean"]):
            mask_dict.update(
                zip([f"{n}_{prop_n}" for n in channel_names_loaded], [x[prop_n] for x in dict_of_chan[nm]])
            )
        # Get the cell IDs and mask properties
        mask_properties = pd.DataFrame(MaskIDs(masks_loaded[nm], mask_props=mask_props))
        mask_dict.update(mask_properties)
        dict_of_chan[nm] = pd.DataFrame(mask_dict).reindex(columns=sorted(mask_dict.keys(), key=col_sort))

    # Return the dict of dataframes for each mask
    return dict_of_chan

def ExtractSingleCells(masks,image,channel_names,output, mask_props=None, intensity_props=["intensity_mean"]):
    """Function for extracting single cell information from input
    path containing single-cell masks, z_stack path, and channel_names path."""

    #Create pathlib object for output
    output = Path(output)

    #Read csv channel names
    channel_names_loaded = pd.read_csv(channel_names)
    #Check for the presence of `marker_name` column
    if 'marker_name' in channel_names_loaded:
        #Get the marker_name column if more than one column (CyCIF structure)
        channel_names_loaded_list = list(channel_names_loaded.marker_name)
    #Consider the old one-marker-per-line plain text format
    elif channel_names_loaded.shape[1] == 1:
        #re-read the csv file and add column name
        channel_names_loaded = pd.read_csv(channel_names, header = None)
        channel_names_loaded_list = list(channel_names_loaded.iloc[:,0])
    else:
        raise Exception('%s must contain the marker_name column'%channel_names)

    #Contrast against the number of markers in the image
    if len(channel_names_loaded_list) != n_channels(image):
        raise Exception("The number of channels in %s doesn't match the image"%channel_names)
    
    #Check for unique marker names -- create new list to store new names
    channel_names_loaded_checked = []
    for idx,val in enumerate(channel_names_loaded_list):
        #Check for unique value
        if channel_names_loaded_list.count(val) > 1:
            #If unique count greater than one, add suffix
            channel_names_loaded_checked.append(val + "_"+ str(channel_names_loaded_list[:idx].count(val) + 1))
        else:
            #Otherwise, leave channel name
            channel_names_loaded_checked.append(val)

    #Read the masks
    masks_loaded = {}
    #iterate through mask paths and read images to add to dictionary object
    for m in masks:
        m_full_name = os.path.basename(m)
        m_name = m_full_name.split('.')[0]
        masks_loaded.update({str(m_name):skimage.io.imread(m,plugin='tifffile')})

    scdata_z = MaskZstack(masks_loaded,image,channel_names_loaded_checked, mask_props=mask_props, intensity_props=intensity_props)
    #Write the singe cell data to a csv file using the image name

    im_full_name = os.path.basename(image)
    im_name = im_full_name.split('.')[0]

    # iterate through each mask and export csv with mask name as suffix
    for k,v in scdata_z.items():
        # export the csv for this mask name
        scdata_z[k].to_csv(
                            str(Path(os.path.join(str(output),
                            str(im_name+"_{}"+".csv").format(k)))),
                            index=False
                            )


def MultiExtractSingleCells(masks,image,channel_names,output, mask_props=None, intensity_props=["intensity_mean"]):
    """Function for iterating over a list of z_stacks and output locations to
    export single-cell data from image masks"""

    print("Extracting single-cell data for "+str(image)+'...')

    #Run the ExtractSingleCells function for this image
    ExtractSingleCells(masks,image,channel_names,output, mask_props=mask_props, intensity_props=intensity_props)

    #Print update
    im_full_name = os.path.basename(image)
    im_name = im_full_name.split('.')[0]
    print("Finished "+str(im_name))