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Segmentation in ImageJ/Fiji

The Basics of Segmentation Methods using ImageJ-based Tools

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What is Segmentation?

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What is Segmentation?

The partitioning of a digital image into multiple segments.

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Common assumptions/requirements:

  1. Do you have overlapping objects? And do you care?

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Common assumptions/requirements:

2. Do you want to count objects? Or do you want to segment regions?

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Common assumptions/requirements:

3. What are the shapes of your objects? Tubes? Blobs? Clouds?

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Common assumptions/requirements:

4. What modality was used? And how does the signal look?

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Segmentation using Machine Learning...

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Segmentation using Machine Learning...

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Segmentation using Machine Learning...

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Segmentation using Machine Learning...

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How to Segment?

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How to Segment?

A Typical Pipeline

1. Preprocess

2. Threshold

3. Cleanup

4. Identify

5. Measure

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How to Segment?

1. Preprocess (Link to sample image)

Improve signal-to-noise ratio

Find Edges | Kuwahara | Gaussian Blur | Median | Background Correction | Deconvolution

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Getting Started with Segmentation

2. Threshold

Separate foreground from background

Global | Local | Auto Threshold

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Getting Started with Segmentation

3. Cleanup

Improve mask with morphological operations

Erode | Dilate | Open | Close | Skeletonize | Outline | Fill Holes | Watershed

MorphoLibJ | Morphological Segmentation | Distance Transform Watershed

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How to Segment?

4. Identify

Split into individual objects

Analyze Particles | Extended Particle Analyzer

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Getting Started with Segmentation

5. Analyze

Measure the original data

Set Measurements | ROI Manager

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How to Segment?

Other techniques:

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Real World Examples

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Real World Examples

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Real World Examples

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Real World Examples

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Real World Examples

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Helpful Resources:

  • Help from the community - Scientific Community Image Forum:

  • ImageJ User Guides:

  • Slides from an excellent Segmentation presentation:

  • Additional workshops and presentations: