1 of 25

Segmentation in ImageJ

The Basics of Segmentation Methods/Tools using ImageJ

2 of 25

What is Segmentation?

3 of 25

What is Segmentation?

The partitioning of a digital image into multiple segments.

4 of 25

Common assumptions/requirements:

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

5 of 25

Common assumptions/requirements:

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

6 of 25

Common assumptions/requirements:

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

7 of 25

Common assumptions/requirements:

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

8 of 25

Machine Learning in Segmentation...

9 of 25

Machine Learning in Segmentation...

10 of 25

Machine Learning in Segmentation...

11 of 25

Machine Learning in Segmentation...

12 of 25

How to Segment?

13 of 25

How to Segment?

A Typical Pipeline

1. Preprocess

2. Threshold

3. Cleanup

4. Identify

5. Measure

14 of 25

How to Segment?

1. Preprocess (Link to sample image)

Improve signal-to-noise ratio

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

15 of 25

How to Segment?

2. Threshold

Separate foreground from background

Global | Local | Available methods

16 of 25

How to Segment?

3. Cleanup

Improve mask with morphological operations

Erode | Dilate | Open | Close | Skeletonize | Outline | Fill Holes | Watershed | MorphoLibJ | Morphological Segmentation | Distance Transform Watershed

17 of 25

How to Segment?

4. Identify

Split into individual objects

Analyze Particles | Extended Particle Analyzer

18 of 25

How to Segment?

5. Analyze

Measure the original data

Set Measurements

19 of 25

How to Segment?

Other techniques:

20 of 25

Real World Examples

21 of 25

Real World Examples

22 of 25

Real World Examples

23 of 25

Real World Examples

24 of 25

Real World Examples

25 of 25

Helpful Resources:

  • Help from the community - Scientific Community Image Forum:

  • ImageJ User Guides:

  • Slides from an excellent Segmentation presentation:

  • Additional workshops and presentations: