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Segmentation of cells based on label-free microscopy

Description

This repository contains a TensorFlow implementation of a segmentation algorithm, a modified U-Net achieving accurate segmentation of various cells in a label-free living tumor microenvironment.

About

Many advances have been made in the area of microscopy thanks to the rapid development of markers and labels. However, marker-based analyses have fundamental limitations due to its perturbance to the living system. Label-free nonlinear optical microscopy, which produces high-resolution images with rich functional and structural information based on intrinsic molecular contrast, has demonstrated strong potential to overcome these problems by generating a broader array of volumetric signals from tissue structures and molecular composition. Here, we shared a multiclass pixel-level neural network program that segments the major components of the tumor microenvironment, including tumor cells, stromal components (fibroblasts, endothelial cells, lymphocytes, red blood cells, adipocytes), and EVs.

Installation

Requirement:

  • TensorFlow >=1.12.0
  • Skimage

Usage

Set directories

  • homedir: home directory
  • logdir: log directory
  • traindir: training directory where the mask files for the training data are saved
  • validationdir: validation directory where files for the validation data were saved
  • testdir: testing direcotry where raw files for testing were saved
  • resultModeldir: save models to this directory
  • resultsImagedir: save validation images to this directory
  • resultsApplydir: save results from applying the model to test images to this directory

Set parameters:

  • ous: network output size
  • ins: network input size (random cropping size during the training)
  • interv: difference of gray value in mask labeling
  • batch_size: batch size for training
  • iterModel: whether to load previous model or to train from scratch
  • iterMax: maximum iterations to run
  • parameters: saved name including all the parameters
  • imageType: how many modalities
  • nc: how many channels

Example

Training

python3 segmentation.py

Testing

python3 segmentation.py -model homedir/resultsModeldir/15000.ckpt -test_dir homedir/testdir\ png/

Citation

If you find this useful or use this method in your work, please cite: Sixian You, Eric J. Chaney, Haohua Tu, Yi Sun, Saurabh Sinha, Stephen A. Boppart. Label-free deep profiling of the tumor microenvironment.

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