live-dead-analysis

Analyse 3D Live/Dead images by an approximation to 2D

https://github.com/deepshika96/live-dead-analysis

Science Score: 57.0%

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Keywords

3d bioprinting organoids viability
Last synced: 6 months ago · JSON representation ·

Repository

Analyse 3D Live/Dead images by an approximation to 2D

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  • Host: GitHub
  • Owner: deepshika96
  • License: mit
  • Default Branch: main
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3d bioprinting organoids viability
Created 9 months ago · Last pushed 9 months ago
Metadata Files
Readme License Citation

README.md

Live-Dead-Analysis

🔬 Quantitative Analysis of DAPI+ Cells and Caspase-3 Activation

This repository contains two complementary scripts designed to analyze confocal microscopy data:

  1. Estimate the average size of DAPI+ cells from randomly sampled patches. - Use Single Cell Area Calculation
  2. Measure DAPI and Caspase-3 (clCASP3) stain coverage and intensity across .lif image series. - Use Stain Area Coverage Analysis
  3. Normalize Caspase-3 intensity by DAPI cell size to get per-cell apoptotic burden.


🧪 1. Estimating Average DAPI+ Cell Area

Script: Single Cell Area Calculation.py

This script: - Loads a .lif file with DAPI-stained nuclei. - Extracts non-overlapping random patches from the DAPI channel. - Segments each patch using the CellSAM deep-learning model. - Counts objects (nuclei) and total DAPI+ area per patch. - Saves masks and an Excel summary.

📤 Output:

  • Labeled TIFF masks per patch
  • Excel file with:
    • Number of DAPI+ cells per patch
    • Total DAPI+ intensity per patch

✅ Final Result:

Use this to compute the average area per DAPI+ nucleus:


🔍 2. DAPI & Caspase-3 Area Coverage

Script: Stain Area Coverage Analysis.py

This script: - Loads a .lif file with DAPI and clCASP3 channels. - Computes max-intensity projections (MIPs). - Extracts 10 random non-overlapping patches per image series. - Applies Otsu thresholding on each channel. - Calculates area (µm²) and intensity of DAPI+ and Caspase-3+ pixels. - Saves overlays and exports a detailed Excel report.

📤 Output:

  • Binary masks for each patch
  • RGB overlay images (R = Caspase-3, B = DAPI)
  • Excel file with:
    • DAPI and Caspase-3 area and intensity per patch

📊 3. Final Step: Normalizing Caspase-3 Intensity

With both Excel files:

  1. From estimate_dapi_cell_area.py: compute the mean DAPI+ cell area.
  2. From stain_area_coverage_analysis.py: use Caspase-3 total intensity per patch.
  3. Normalize clCASP3 intensity per cell:

CellSAM is used in this code, find more here: @article{israel2023foundation, title={A Foundation Model for Cell Segmentation}, author={Israel, Uriah and Marks, Markus and Dilip, Rohit and Li, Qilin and Schwartz, Morgan and Pradhan, Elora and Pao, Edward and Li, Shenyi and Pearson-Goulart, Alexander and Perona, Pietro and others}, journal={bioRxiv}, publisher={Cold Spring Harbor Laboratory Preprints}, doi = {10.1101/2023.11.17.567630}, }

Owner

  • Login: deepshika96
  • Kind: user

Citation (citation.cff)

cff-version: 1.2.0
title: LiveDeadAnalysis
message: >-
  If you use this software, please cite it using the
  metadata from this file.
type: software
authors:
  - given-names: Deepshika
    family-names: Arasu
    email: deepshika96@gmail.com
    affiliation: University Of Barcelona
identifiers:
  - type: url
    value: 'https://github.com/deepshika96/Live-Dead-Analysis'
abstract: >-
  Performs Live Dead analysis on 3D lif images-
  approximation treats images as 2D.
license: MIT-0

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Dependencies

requirements.txt pypi
  • cellSAM *
  • ipywidgets *
  • matplotlib *
  • numpy *
  • pandas *
  • readlif *
  • scikit-image *
  • tifffile *