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Automatic
Cell Counting
Ege Engin
Middle East Technical University
Supervisor: Tomas Lukes
Czech Technical University
http:/blog.metu.edu.tr/e174088
Agenda
• Introduction
• Different Approaches
• Method 1
• Method 2
• Method 3

• Comparison & Results & Comments
• Nine-Point Circle for Cell Segmentation
•
•
•
•

Nine-Point Circle Rule
How to use?
Results
Implementation

• Conclusion and further work
Background
• Learn image processing basics
•
•
•
•
•

Fundamentals
Intensity Transforms
Spatial Filtering
Frequency Domain Processing
Image Segmentation

• Implement different methods in MATLAB and compare them
• Based on the results, improve the algorithm
1st and 2nd Method
1st Method

• Thresholding:
• im2bw with graythresh

• Filling with holes
• imfill with holes

• Counting:
• Numobjects of
bwconncomp

2nd Method

• Thresholding:
• im2bw with graythresh

• Filling with holes
• imfill with holes

• Counting:
• Lenght of bwboundaries
3rd Method
3rd Method
• Image: Cell 1
Comparison & Results
1st
2nd
3rd
3rd Method
Method Method Method ( with watershed)

Exact Number
(includes incomplete
objects)

Coins

10

37

10

12

10

Eight

1

263

4

3

4

Rice

151

158

93

67

101

Cell Image 1

36

518

53

42

50

Cell Image 2

270

2369

50

44

38

Cell Image 3

287

1139

25

75

64

• Cell Image 1

• Cell Image 2

• Cell Image 3
Comments
• First two approaches uses nearly the same algorithm except their
counting methods.
• The first algorithm underestimate the number of objects.
• The second algorithm overestimate the number of objects.

• 3rd approach :
• Without watershed segmentation has achieved promising results
when the cells in the image are not connected.
• With watershed segmentation over-segments or mis-segments the
figure, so overestimate the number of objects.

• Challenges: connected cells, incomplete cells on the borders
• Another solution technique can be useful for the solution of
connected cells problem.
Nine-Point Circle
• Also known as the Feuerbach Circle

• In every triangle,
lie on a circle:
• The three midpoints of the sides
• The three base points(feet) of the altitudes
• The midpoints of the three segments from the orthocenter to the
vertices
Reference: Dorrie, H. and Woltermann, M, '100 Great Problems of Elementary Mathematics', reworked in 2010
How to use?
• Arbitrarily select three points from cell
• Example: A,B,C

• Find 9 points which are
• 3 midpoints
• Example: D,E,F

• 3 base points(feet) of the altitudes
• Example: G,H,I

• Midpoints of 3 segments
from the orthocenter to the vertices
• Example: K,L,M
• Please note that: J is orthocenter.

• Count the points inside the cell:
• D,E,G,K,L,M => 6 inside / 9 total

• Repeat the procedure until the average is an appropriate result
Results
• For Results:

• For Implementation:
Conclusion
• When useful?
• When cells have circular shapes
• Why?
• The more circular the shape, higher the average algorithm gives
so,When the cells are not connected, the average will be higher.
• Can help to distinguish the cells whether connected or not.

• Why important?
• For cell segmentation, not tried before ( based on subjective(my) research)

• Challanges?
• Defining the standard to understand whether cells are connected
• Clear bordering is necessary to run correctly
• Hard to distinguish if cells are connected circularly
Future Study
• For nine-point rule cell segmentation:
• Divide the given image in appropriate divisions
• Try on cell images

• Documentation related to all studies
Questions

Reference: Velasquez J., Dhuru P. & Vaghani M. , ‘Matlab Digital Image Processing
For Microscopy Screening’
Thank you!

Reference: A blood test and examination, 1941 – 1945 retrieved by
http://research.archives.gov/description/513715 on 20.08.2013
3rd Method
3rd Method
3rd Method

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Ege Engin | Automatic Cell Counting Presentation

  • 1. Automatic Cell Counting Ege Engin Middle East Technical University Supervisor: Tomas Lukes Czech Technical University http:/blog.metu.edu.tr/e174088
  • 2. Agenda • Introduction • Different Approaches • Method 1 • Method 2 • Method 3 • Comparison & Results & Comments • Nine-Point Circle for Cell Segmentation • • • • Nine-Point Circle Rule How to use? Results Implementation • Conclusion and further work
  • 3. Background • Learn image processing basics • • • • • Fundamentals Intensity Transforms Spatial Filtering Frequency Domain Processing Image Segmentation • Implement different methods in MATLAB and compare them • Based on the results, improve the algorithm
  • 4. 1st and 2nd Method 1st Method • Thresholding: • im2bw with graythresh • Filling with holes • imfill with holes • Counting: • Numobjects of bwconncomp 2nd Method • Thresholding: • im2bw with graythresh • Filling with holes • imfill with holes • Counting: • Lenght of bwboundaries
  • 7. Comparison & Results 1st 2nd 3rd 3rd Method Method Method Method ( with watershed) Exact Number (includes incomplete objects) Coins 10 37 10 12 10 Eight 1 263 4 3 4 Rice 151 158 93 67 101 Cell Image 1 36 518 53 42 50 Cell Image 2 270 2369 50 44 38 Cell Image 3 287 1139 25 75 64 • Cell Image 1 • Cell Image 2 • Cell Image 3
  • 8. Comments • First two approaches uses nearly the same algorithm except their counting methods. • The first algorithm underestimate the number of objects. • The second algorithm overestimate the number of objects. • 3rd approach : • Without watershed segmentation has achieved promising results when the cells in the image are not connected. • With watershed segmentation over-segments or mis-segments the figure, so overestimate the number of objects. • Challenges: connected cells, incomplete cells on the borders • Another solution technique can be useful for the solution of connected cells problem.
  • 9. Nine-Point Circle • Also known as the Feuerbach Circle • In every triangle, lie on a circle: • The three midpoints of the sides • The three base points(feet) of the altitudes • The midpoints of the three segments from the orthocenter to the vertices Reference: Dorrie, H. and Woltermann, M, '100 Great Problems of Elementary Mathematics', reworked in 2010
  • 10. How to use? • Arbitrarily select three points from cell • Example: A,B,C • Find 9 points which are • 3 midpoints • Example: D,E,F • 3 base points(feet) of the altitudes • Example: G,H,I • Midpoints of 3 segments from the orthocenter to the vertices • Example: K,L,M • Please note that: J is orthocenter. • Count the points inside the cell: • D,E,G,K,L,M => 6 inside / 9 total • Repeat the procedure until the average is an appropriate result
  • 11. Results • For Results: • For Implementation:
  • 12. Conclusion • When useful? • When cells have circular shapes • Why? • The more circular the shape, higher the average algorithm gives so,When the cells are not connected, the average will be higher. • Can help to distinguish the cells whether connected or not. • Why important? • For cell segmentation, not tried before ( based on subjective(my) research) • Challanges? • Defining the standard to understand whether cells are connected • Clear bordering is necessary to run correctly • Hard to distinguish if cells are connected circularly
  • 13. Future Study • For nine-point rule cell segmentation: • Divide the given image in appropriate divisions • Try on cell images • Documentation related to all studies
  • 14. Questions Reference: Velasquez J., Dhuru P. & Vaghani M. , ‘Matlab Digital Image Processing For Microscopy Screening’
  • 15. Thank you! Reference: A blood test and examination, 1941 – 1945 retrieved by http://research.archives.gov/description/513715 on 20.08.2013