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DOI

Vectra Imaging Processing Pipeline

Multiplexed imaging data is providing dramatic opportunities to understand the tumor microenvironment, but there is an acute need for better analysis tools. Here, we provide a pipeline for multiplexed imaging quality control and processing. It contains three core steps:

  1. Preprocess raw images to remove undesired noise (introduced by technical sources) while retaining biological signal
  2. Perform segmentation to draw boundaries around individual cells, making it possible to discern morphology and which features, such as detected RNA or protein, belong to each cell.
  3. Extract cellular feature from images via segmentation and assign cell types to each cell.

To transform digital images into cell-level measurements, we have been applying, comparing and optimizing cutting-edge computer vision and machine learning techniques to each of the steps above. All code is written in Python.

Quick Start

Dependencies

  • Clone this repository

  • Create a new conda environment for this pipeline

  • conda env create -n davinci -f environment.yml 
    source activate davinci
    python -m ipykernel install --user --name davinci
  • Install this package

  • python setup.py install
  • Install the deep learning model for segmentation following the instruction on link

Usage

Step-by-step tutorial on the usage can be found in the following Jupiter notebooks.

More details can be found in the technical notes

0. Data inspection

1. Preprocessing

2. Segmentation

  • Train deep learning model on custom data Notebook
  • Predict segmentation with pre-trained model Notebook

3. Cell typing

Workflow

img

Future plan

  • Provide more pre-trained weights for segmentation using different marker panels

Note

  • The data in this repo are for demonstration only. Please do not use them for any other purposes.

Acknowledgement

This work is supported by

Reference

He, K., G. Gkioxari, P. Dollár, and R. Girshick. 2017. “Mask R-CNN.” In 2017 IEEE International Conference on Computer Vision (ICCV), 2980–88. link