Quantification of Ki-67 labeling index in pediatric brain tumor immunohistochemistry images.
The 2 matches
- [1] § RESULTS › Density maps ↔ summary_ratios.py, lines 106–146 · score 0.61 · QuPath, LI map, positive cell density, negative cell density, ratio, slide
- [2] § METHODS › Postprocessing in Python ↔ summary_ratios.py, lines 106–146 · score 0.60 · QuPath, LI map, cell density maps, ratio, negative cell, Ki
Paper
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The authors' code
Python · 389 lines · 16 KB · GPL-3.0 · 2 matches
- # %% IMPORTS
- import json
- import os
- import numpy as np
- import math
- import pandas as pd
- import argparse
- import matplotlib
- import matplotlib.pyplot as plt
- import seaborn as sns
- from PIL import Image
- from mpl_toolkits.axes_grid1 import make_axes_locatable
- # %% ARGUMENT PARSING
- parser = argparse.ArgumentParser(description='Create summary ratios and density maps.')
- parser.add_argument('--maps_dir', type=str, default=False, help='Directory of the cell density maps generated by QuPath.')
- parser.add_argument('--data_dir', type=str, default=False, help='Directory of the project data')
- parser.add_argument('--area_path', type=str, default=False, help='Path to Area.txt which is saved under the Results folder.')
- parser.add_argument('--csv_path', type=str, required=True, help='Path to the WSI dataframe CSV file')
- parser.add_argument('--WSIs_dir', type=str, default=False, help='Directory of the folder where the WSIs exist.')
- parser.add_argument('--norm_maps_dir', type=str, default=False, help='Directory to save the normalized cell density maps.')
- parser.add_argument('--result_dir', type=str, default=False, help='Directory to save the summary tables and graphs.')
- args = parser.parse_args()
- # %% DIRECTORIES & FILES NEEDED FOR THE ANALYSIS
- maps_dir = args.maps_dir
- data_dir = args.data_dir
- csv_path = args.csv_path
- # check if the CSV file exists
- if not os.path.isfile(csv_path):
- raise FileNotFoundError(f"The specified CSV file does not exist: {csv_path}")
- area_path = args.area_path
- # check if the Area.txt file exists
- if not os.path.isfile(area_path):
- raise FileNotFoundError(f"The specified Area.txt file does not exist: {area_path}")
- WSI_df = pd.read_csv(csv_path)
- WSIs_dir = args.WSIs_dir
- norm_maps_dir = args.norm_maps_dir
- # create the directory to save the normalised maps if it does not exist
- if not os.path.exists(norm_maps_dir):
- os.makedirs(norm_maps_dir)
- result_dir = args.result_dir
- # create the directory to save the results if it does not exist
- if not os.path.exists(result_dir):
- os.makedirs(result_dir)
- # get the area and total detections
- annotation_df = pd.read_csv(area_path,names=['name', 'detections', 'area'],delimiter=";",index_col=False)
- # %% RETRIEVE INFORMATION FROM JSON FILES
- file_list = [f for f in os.listdir(data_dir) if os.path.isdir(os.path.join(data_dir,f))]
- WSIs = os.listdir(WSIs_dir)
- # data frame to store the results
- results_df = pd.DataFrame(columns = ['case_id', 'slide_id', 'label', 'Positive_Cell_Count', 'Positive_Cell_Density', 'Negative_Cell_Count', 'Negative_Cell_Density', 'Ki-67 LI', 'Cell_Density'])
- proc_l = len(file_list)
- proc = -1
- failed_list = []
- failed_ann = []
- for folder in file_list:
- proc += 1
- proc_perc = round(proc/proc_l*100, 2)
- print('Progress: ' + str(proc_perc) + '%')
- print('Processing image ' + str(proc) + ' of ' + str(proc_l))
- with open(data_dir + '/' + folder + '/server.json', 'r') as s_file:
- server_data = json.load(s_file)
- slide_id = server_data['builder']['metadata']['name'][:-4]
- case_id = slide_id.split('___')[0]
- # Convert slide_id to int to match CSV data type
- try:
- slide_id_lookup = int(slide_id)
- except ValueError:
- # If conversion fails, keep as string (for cases with non-numeric IDs)
- slide_id_lookup = slide_id
- label = WSI_df['label'].loc[WSI_df['slide_id']==slide_id_lookup].values[0]
- print('Reading data from image ' + slide_id + ':')
- with open(data_dir + '/' + folder + '/summary.json', 'r') as file:
- data = json.load(file)
- detection_data = data['hierarchy']['detectionClassificationCounts']
- if 'Positive' not in detection_data.keys() and 'Negative' not in detection_data.keys():
- print('The slide has not been properly segmented yet, skipping.')
- failed_list.append(slide_id)
- continue
- # calculate the Ki-67 LI
- pos_cell_count = detection_data.get('Positive', 0)
- neg_cell_count = detection_data.get('Negative', 0)
- # Check if there are any cells detected
- if pos_cell_count + neg_cell_count == 0:
- print('No cells detected in this slide, skipping.')
- failed_list.append(slide_id)
- continue
- # round ratio to 2 decimal places
- ki67_li = round(pos_cell_count / (pos_cell_count + neg_cell_count), 2)
- for WSI in WSIs:
- WSI_name = WSI.split('.')[0]
- if slide_id in WSI_name:
- slide_file_name = WSI
- print('Found the corresponding WSI file: ' + slide_file_name)
- break
- try:
- annAreamm = annotation_df['area'].loc[annotation_df['name'] == slide_file_name].values[0]*0.000001
- numdet = annotation_df['detections'].loc[annotation_df['name'] == slide_file_name].values[0]
- cells_smm = numdet/annAreamm
- pos_cell_dens = pos_cell_count/annAreamm
- neg_cell_dens = neg_cell_count/annAreamm
- except:
- print('Could not find any annotation data.')
- failed_ann.append(slide_file_name)
- else:
- print("Area of the segmented tissue: " + str(annAreamm))
- print("Total number of detections: " + str(numdet))
- print('Number of detections per mm^2: ' + str(cells_smm))
- results_df.loc[len(results_df)] = [case_id, slide_id, label, pos_cell_count, pos_cell_dens, neg_cell_count, neg_cell_dens, ki67_li*100, cells_smm]
- print('Positive cell count: ' + str(pos_cell_count))
- print('Negative cell count: ' + str(neg_cell_count))
- print('Positive to negative ratio: ' + str(ki67_li))
- # %% GENERATE THE NORMALISED AND RATIO CELL DENSITY MAPS
- # get the cell density maps generated by QuPath
- HM_file_neg = maps_dir + '/' + slide_id + '_NegDMap.tif'
- HM_file_pos = maps_dir + '/' + slide_id + '_PosDMap.tif'
- failed_cell_density_list = []
- # check if both cell density map files exist before proceeding
- if os.path.isfile(HM_file_neg) and os.path.isfile(HM_file_pos):
- neg_pp_im = norm_maps_dir + '/' + slide_id + '_Neg_Cell_Dens_Map.png'
- pos_pp_im = norm_maps_dir + '/' + slide_id + '_Pos_Cell_Dens_Map.png'
- ratio_pp_im = norm_maps_dir + '/' + slide_id + '_Ki-67_LI_Map.png'
- # generate the normalised negative cell density map
- if not os.path.isfile(neg_pp_im):
- print("Generating normalized negative cell density map.")
- # get the raw grayscale negative cell density map
- im_neg = Image.open(HM_file_neg)
- width, height = im_neg.size
- # set color bar orientation depending on image dimensions:
- if width < height:
- orient = 'vertical'
- else:
- orient = 'horizontal'
- # normalize
- imarray_neg = np.array(im_neg)
- cm_lim_neg = np.max(imarray_neg)
- new_imarray_neg = imarray_neg/cm_lim_neg
- # keep values inside normalized range for a correct comparison between heat maps
- new_imarray_neg[new_imarray_neg>1] = 1
- # colorise the negative cell density map
- cmapQ = plt.get_cmap('jet')
- A_neg = cmapQ(new_imarray_neg)
- # convert dark blue background to white
- A_neg[new_imarray_neg == 0] = [1, 1, 1, 1]
- # plot and save normalised negative cell density map
- fig, ax = plt.subplots()
- im = ax.imshow(A_neg)
- ax.set_xticks([])
- ax.set_yticks([])
- ax.set_title('Negative Cell Density Map')
- divider = make_axes_locatable(ax)
- if orient == 'vertical':
- cax = divider.append_axes("right", size="5%", pad=0.20)
- else:
- cax = divider.append_axes("bottom", size="5%", pad=0.20)
- norm = matplotlib.colors.Normalize(vmin=0, vmax=neg_cell_dens)
- cb = plt.colorbar(plt.cm.ScalarMappable(norm=norm, cmap='jet'), cax=cax, orientation=orient, shrink=1.0)
- plt.title('Negative Cell Density Map')
- plt.savefig(neg_pp_im)
- cb.remove()
- plt.close("all")
- # mask of the negative cell density map
- A_neg_blue_mask = A_neg
- A_neg_blue_mask[np.any(A_neg_blue_mask[:, :, :3] != [1, 1, 1], axis=2)] = [0, 0, 1, 1]
- else:
- print("Normalized negative cell density map already exists, skipping.")
- # generate the normalised positive cell density map
- if not os.path.isfile(pos_pp_im):
- print("Generating normalized positive cell density map.")
- # get the raw grayscale positive cell density map
- im_pos = Image.open(HM_file_pos)
- width, height = im_pos.size
- # set color bar orientation depending on image dimensions:
- if width < height:
- orient = 'vertical'
- else:
- orient = 'horizontal'
- # normalise
- imarray_pos = np.array(im_pos)
- cm_lim_pos = np.max(imarray_pos)
- new_imarray_pos = imarray_pos/cm_lim_pos
- # keep values inside normalized range for a correct comparison between heat maps
- new_imarray_pos[new_imarray_pos>1] = 1
- # colorise
- cmapQ = plt.get_cmap('jet')
- A_pos = cmapQ(new_imarray_pos)
- # convert dark blue background to white
- A_pos[new_imarray_pos == 0] = [1, 1, 1, 1]
- new_A_pos = A_pos
- white_mask = np.all(new_A_pos[:, :, :3] == [1, 1, 1], axis=2)
- new_A_pos[white_mask] = A_neg_blue_mask[white_mask]
- # plot and save normalised positive cell density map
- fig, ax = plt.subplots()
- im = ax.imshow(new_A_pos)
- ax.set_xticks([])
- ax.set_yticks([])
- ax.set_title('Positive Cell Density Map')
- divider = make_axes_locatable(ax)
- if orient == 'vertical':
- cax = divider.append_axes("right", size="5%", pad=0.20)
- else:
- cax = divider.append_axes("bottom", size="5%", pad=0.20)
- norm = matplotlib.colors.Normalize(vmin=0, vmax=pos_cell_dens)
- cb = plt.colorbar(plt.cm.ScalarMappable(norm=norm, cmap='jet'), cax=cax, orientation=orient, shrink=1.0)
- plt.savefig(pos_pp_im)
- cb.remove()
- plt.close("all")
- else:
- print("Normalized positive cell density map already exists, skipping.")
- # generate the Ki-67 LI map
- if not os.path.isfile(ratio_pp_im):
- print("Generating ratio cell density map.")
- neg_im = Image.open(HM_file_neg)
- width, height = neg_im.size
- # set color bar orientation depending on image dimensions:
- if width < height:
- orient = 'vertical'
- else:
- orient = 'horizontal'
- neg_imarray = np.array(neg_im)
- pos_im = Image.open(HM_file_pos)
- pos_imarray = np.array(pos_im)
- for k in range(np.shape(neg_imarray)[0]):
- for l in range(np.shape(neg_imarray)[1]):
- denominator = neg_imarray[k, l] + pos_imarray[k, l]
- # if denominator is 0, assign a default value
- if denominator == 0:
- neg_imarray[k, l] = 0.0001
- else:
- neg_imarray[k, l] = pos_imarray[k, l] / denominator
- if neg_imarray[k, l] < 0.0001:
- neg_imarray[k, l] = 0.0001
- elif neg_imarray[k, l] > 1:
- neg_imarray[k, l] = 1
- elif math.isnan(neg_imarray[k, l]):
- neg_imarray[k, l] = 0.0001
- # only take into account foreground pixels
- relevant_values = neg_imarray[neg_imarray > 0]
- n_relval = len(relevant_values)
- adjusted_n_relval = round(n_relval*0.95)
- sorted_relval = sorted(relevant_values)
- min_cm_lim = 0.03
- cm_lim = max(max(sorted_relval[:adjusted_n_relval]), min_cm_lim)
- # normalize with obtained value
- neg_imarray = neg_imarray/cm_lim
- # keep values inside normalized range for a correct comparison between heat maps
- neg_imarray[neg_imarray>1] = 1
- # colorise
- cmapQ = plt.get_cmap('jet')
- A_ki67_li = cmapQ(neg_imarray)
- # convert dark blue background to white
- dark_blue = [0, 0, 0.5, 1] # RGBA values for dark blue
- white = [1, 1, 1, 1] # RGBA values for white
- A_ki67_li[
- (A_ki67_li[:, :, 0] == dark_blue[0]) &
- (A_ki67_li[:, :, 1] == dark_blue[1]) &
- (A_ki67_li[:, :, 2] == dark_blue[2]) &
- (A_ki67_li[:, :, 3] == dark_blue[3])
- ] = white
- new_A_ki67_li = A_ki67_li
- white_mask = np.all(new_A_ki67_li[:, :, :3] == [1, 1, 1], axis=2)
- new_A_ki67_li[white_mask] = A_neg_blue_mask[white_mask]
- # plot and save the ki-67 li map
- fig, ax = plt.subplots()
- im = ax.imshow(new_A_ki67_li)
- ax.set_xticks([])
- ax.set_yticks([])
- ax.set_title('Ki-67 LI Map')
- divider = make_axes_locatable(ax)
- if orient == 'vertical':
- cax = divider.append_axes("right", size="5%", pad=0.20)
- else:
- cax = divider.append_axes("bottom", size="5%", pad=0.20)
- norm = matplotlib.colors.Normalize(vmin=0, vmax=ki67_li)
- cb = plt.colorbar(plt.cm.ScalarMappable(norm=norm, cmap='jet'), cax=cax, orientation=orient, shrink=1.0)
- plt.savefig(ratio_pp_im)
- cb.remove()
- plt.close("all")
- else:
- print("Normalized Ki-67 LI map already exists, skipping.")
- else:
- print(f"Required cell density map files not found for slide ID: {slide_id}. Skipping.")
- # %% CREATE SUMMARY STATISTICS
- # save the results to a csv file
- results_df.to_csv(os.path.join(result_dir, 'summary_results.csv'), index=False)
- statistics_df = pd.DataFrame(columns = ['variable', 'label', 'mean', 'sd', 'median', 'minimum', 'maximum'])
- summary_points = ['Positive_Cell_Count', 'Positive_Cell_Density', 'Negative_Cell_Count', 'Negative_Cell_Density', 'Ki-67 LI', 'Cell_Density']
- y_units = {'Positive_Cell_Count': 'Number of positive cells',
- 'Positive_Cell_Density': 'Number of positive cells per mm^2',
- 'Negative_Cell_Count': 'Number of negative cells',
- 'Negative_Cell_Density': 'Number of negative cells per mm^2',
- 'Ki-67 LI': 'Ki-67 LI',
- 'Cell_Density': 'Number of cells per mm^2'}
- res_df = pd.read_csv(os.path.join(result_dir, 'summary_results.csv'))
- label_list = np.unique(res_df['label'].tolist())
- # summary table and boxplots
- for sp in summary_points:
- box_data = []
- for label in label_list:
- sp_array = res_df[sp].loc[res_df['label'] == label]
- sp_array = sp_array[~np.isnan(sp_array)]
- box_data.append(sp_array)
- sp_mean = round(np.mean(sp_array), 2)
- sp_sd = round(np.std(sp_array), 2)
- sp_median = round(np.median(sp_array), 2)
- sp_min = round(np.min(sp_array), 2)
- sp_max = round(np.max(sp_array), 2)
- # summary statistics
- statistics_df.loc[len(statistics_df)] = [sp, label, sp_mean, sp_sd, sp_median, sp_min, sp_max]
- # boxplots
- palette = sns.color_palette("deep", 10)
- custom_colors = [palette[9]]
- palette = sns.color_palette(custom_colors * len(label_list))
- sns.set_style("whitegrid", {'axes.grid': False})
- plt.figure(figsize=(10, 6))
- plt.gca().set_facecolor('white')
- flierprops = dict(marker='D', markerfacecolor='darkgrey', markersize=5, linestyle='none')
- boxplot = sns.boxplot(
- x='label',
- y=sp,
- data=res_df,
- flierprops=flierprops,
- palette=palette,
- linewidth=2
- )
- plt.title(f'{sp} across all diagnoses')
- plt.xlabel('Diagnosis')
- plt.ylabel(y_units[sp])
- plt.tight_layout()
- plt.savefig(os.path.join(result_dir,sp + '_BoxPlot.png'))
- plt.close()
- # summary statistics
- statistics_df.to_csv(os.path.join(result_dir, 'summary_statistics.csv'), index=False)
- # %%
summary_ratios.py at commit d1efa7c, under GPL-3.0 · at the source
Overview
- Department of Biomedical Engineering, Linköping University, Linköping, Sweden
- Center for Medical Image Science and Visualization, Linköping University, Linköping, Sweden
- Clinical Department of Clinical Pathology, Region Östergötland, Linköping, Sweden
- Department of Biomedical and Clinical Sciences, Linköping University, Linköping, Sweden
- Crown Princess Victoria Children's Hospital, Region Östergötland, Linköping, Sweden
- Department of Health, Medicine and Caring Sciences, Linköping University, Linköping, Sweden
- Clinical Department of Radiology in Linköping, Region Östergötland, Linköping, Sweden
- Department of Clinical Pathology and Cancer Diagnostics, Karolinska University Hospital, Solna, Sweden
- Department of Oncology-Pathology, Karolinska Institute, Solna, Sweden
- Clinical Department of Oncology in Linköping, Region Östergötland, Linköping, Sweden
Abstract
Quantification of the Kiel 67 (Ki-67) labeling index (LI) is critical for assessing proliferation and prognosis in tumors but manual scoring remains a common practice. We present an automated framework for Ki-67 scoring in whole slide images (WSIs) developed for research settings using an Apache Groovy code script for QuPath and complemented by a Python postprocessing script that provides cell density maps and summary tables. Tissue segmentation is performed by pixel classifiers and cell segmentation is conducted using StarDist, a deep learning model, followed by adaptive thresholding to classify Ki-67 positive and negative nuclei. The pipeline was applied to a cohort of 632 pediatric brain tumor cases with 734 Ki-67 WSIs from the Children’s Brain Tumor Network. Medulloblastomas showed the highest Ki-67 LI (median: 19.84), followed by atypical teratoid rhabdoid tumors (median: 19.36), brainstem glioma-diffuse intrinsic pontine gliomas (median: 11.50), high-grade gliomas (grades 3, 4) (median: 9.50), and ependymomas (median: 5.88). Lower indices were found in meningiomas (median: 1.84) and the lowest were seen in low-grade gliomas (grades 1, 2) (median: 0.85), dysembryoplastic neuroepithelial tumors (median: 0.63), and gangliogliomas (median: 0.50). The results demonstrate a significant correlation (P < .05) in Ki-67 LI across most of the tumor families/
Reproduced under the paper's license (CC BY), from the paper cited above.
Repository
Its files are read in the Code ↔ Paper reader above, with 2 matches between paragraphs and lines of code.
Christoforos-Spyretos/QuPath-Automatic-Cell-Detection-for-Ki-67-WSIs
d1efa7cff94c115107a0e4d9790d606b0eba0134, 21 May 2026Availability: 1 check, the latest on 28 September 2026: the link answers
- 28 September 2026: the link answers
3 files
- summary_ratios.py — Python, 389 lines, 2 matches
- LICENSE — License, 674 lines
- README.md — Text, 195 lines
The paper's code and data availability statement is in the Data section.
Tracing map
Proposed by the machine: these links were found in the paper and verified at the source, without human review. The map will receive a Zenodo DOI once one of the paper's authors has validated it with their ORCID.
What the map holds:
- 1 repository of the authors' code, each at its verified commit, with its license and how the link was found in the paper;
- 1 script, each with its path and the digest of its content;
- 2 matches between paragraphs of the paper and lines of the code (method lexical-v1);
- neither the text of the paper nor the code itself.
Its JSON (tracing-map.json) is deposited on Zenodo with its DOI once the map is validated.
Data
Datasets cited
- github.com/
cialab/ — at github.com; found in “DATA AVAILABILITY”neuroendocrine_ - zenodo:11218961 — at Zenodo; found in “DATA AVAILABILITY”
Data availability
The open-access dataset used in this study was obtained from the Children’s Brain Tumor Network (https://
Reproduced under the paper's license (CC BY), from the paper cited above.
Versions
The history of this record: each version stored by the harvester or made by a correction of its authors or of the maintainers of its code, and what changed in its facts. The texts of the paper (its abstract, its availability statements) are not part of it; versions that changed only those are not listed.
Version 1, 28 September 2026: the first record
Recorded: type, language, journal, volume, issue, pages, dates, 8 authors, 9 keywords, 11 MeSH terms, 8 funders, 29 references.
Cite
This paper
Spyretos, C., Pardo Ladino, J. M., Andersen Blomstrand, H., Nyman, P., Snödahl, O., Shamikh, A., Elander, N., & Haj-Hosseini, N. (2026). Quantification of Ki-67 labeling index in pediatric brain tumor immunohistochemistry images. Journal of neuropathology and experimental neurology, 85(5), 475-486. https://
BibTeX
@article{spyretos2026qua
author = {Spyretos, Christoforos and Pardo Ladino, Juan Manuel and Andersen Blomstrand, Hakon and Nyman, Per and Snödahl, Oscar and Shamikh, Alia and Elander, Nils and Haj-Hosseini, Neda},
title = {{Quantification of Ki-67 labeling index in pediatric brain tumor immunohistochemistry images}},
journal = {Journal of neuropathology and experimental neurology},
year = {2026},
month = may,
volume = {85},
number = {5},
pages = {475--486},
publisher = {Oxford University Press},
issn = {0022-3069},
doi = {10.1093/
url = {https://
pmid = {41806389},
pmcid = {PMC13127889}
}
RIS
TY - JOUR
AU - Spyretos, Christoforos
AU - Pardo Ladino, Juan Manuel
AU - Andersen Blomstrand, Hakon
AU - Nyman, Per
AU - Snödahl, Oscar
AU - Shamikh, Alia
AU - Elander, Nils
AU - Haj-Hosseini, Neda
TI - Quantification of Ki-67 labeling index in pediatric brain tumor immunohistochemistry images
T2 - Journal of neuropathology and experimental neurology
J2 - J Neuropathol Exp Neurol
PY - 2026
DA - 2026/
VL - 85
IS - 5
SP - 475
EP - 486
SN - 0022-3069
PB - Oxford University Press
DO - 10.1093/
UR - https://
LA - en
ER -
CSL-JSON
{
"id": "10.1093/
"type": "article-journal",
"title": "Quantification of Ki-67 labeling index in pediatric brain tumor immunohistochemistry images",
"container-title": "Journal of neuropathology and experimental neurology",
"author": [
{
"family": "Spyretos",
"given": "Christoforos"
},
{
"family": "Pardo Ladino",
"given": "Juan Manuel"
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{
"family": "Andersen Blomstrand",
"given": "Hakon"
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{
"family": "Nyman",
"given": "Per"
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{
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{
"family": "Elander",
"given": "Nils"
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{
"family": "Haj-Hosseini",
"given": "Neda"
}
],
"container-title-short":
"volume": "85",
"issue": "5",
"page": "475-486",
"DOI": "10.1093/
"PMID": "41806389",
"PMCID": "PMC13127889",
"ISSN": "0022-3069",
"publisher": "Oxford University Press",
"URL": "https://
"language": "en",
"issued": {
"date-parts": [
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]
}
}
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