SICE Annual Conference 2011

September 13-18, 2011, Waseda University, Tokyo, Japan

Extraction Method of Glandular Areas in Prostate Biopsy Image

Yoshiteru Toki1 and Toshiyuki Tanaka2
1,2

School of Science and Technology, Keio University, Yokohama, Japan

E-mail: toki@isp.appi.keio.ac.jp

Abstract: Number of prostate cancer patients is increasing. So system of supporting diagnosis is needed by pathologists.
Biopsy images stained by HE staining, but the state depends on pathologist, individual condition, and environments. In
this study, we try to robust image segmentation method to divide glandular into components. Our proposed method is to
use xed color range for background, Otsu method with Laplacian histosgram for nuclei, and not only color information
but also texture features for interstitium and cytoplasm.
Keywords: Image segmentation, texture analysis

1. INTRODUCTION

ing method. In tissue slice, HE stains nuclei to darkpurple, interstitium to magenta, and cytoplasm to lightpurple. But the staining colors arent xed because of
individual difference of patient and etc. So robust imagesegmentation method is needed.

Prostate biopsy is important examination to choice of

therapy for prostate cancer. But the work is heavy because patients number is increasing and the examination is needed to check the samples from end to end. So
reducing the burden is needed. Moreover, there is case
the examination result isnt stable because the diagnostic
standard is qualitative, quantitative diagnostic standard is
needed. Severity determination by biopsy image has been
attempted in medical image engineering. However individual deference of their images is large so automatic
image analysis is hard. The purpose of this study is to
extract glandular areas for calculation severity of prostate
cancer. Shape, size and distribution of glandular areas
reect severity of prostate cancer. For this reason extracting glandular area accurately is important. This study is
trying to divide components of prostate glandular (nuclei,
interstitium, and cytoplasm). Example of prostate biopsy
image is shown on Fig. 1

(a) Original image

(b) Interstitium area

(c) Cytoplasm area

(d) Nuclei area

Fig. 2 Components of glandular

2.1 Dividing components by supervised learning
method
Color space is divided into 4 components area by discriminant analysis. Then training data is made by extracting components from biopsy image manually. A training
data is shown on Fig. 3
First, no data area whose color is (R, G, B) =
(255, 255, 255) was removed on each images. Second,
pixels of components were converted from RGB color
to HSV color. Third, histogram of saturation and value
was made of these pixels each component. Fourth, color
space is dividing into 4 components by discriminant anal-

Fig. 1 A prostate biopsy image.

2. METHOD
A biopsy image is shown on Fig. 1. Biopsy sample is processed by HE (Haematoxylin - Eosin) stain-

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PR0001/11/0000-1995 400 2011 SICE

2.2.1 Background extraction by xed color range

In this study, biopsy image is adjusted to same background color. So background areas could be extracted by
xed color range from our test images. After an image is
binarized and labeled, label of little areas are removed as
noises. In this label image, labels on the image edge are
extracted as background, and others labels are extracted
as glandular inner area.

(c) Nuclei

(d) Background

2.2.3 Interstitium and cytoplasm extraction by textures

Extracting interstitium and cytoplasm is difcult because their difference of color is too little. But, the texture is different on these areas. Therefore we try to divide
these areas with their textures. Interstitium has smooth
textue along glandular contour. On the other hand, cytoprasm has rough texture with cell membrane. First,
edge intensity is calculated by applying Laplacian lter to
biopsy image. We assumed interstitium has low edge intensity and cytoplasm has high edge intensity. After low
and high edge intensity area is extracted, these picels are
grouped by k-means clustering method. By these pixels
groups as learning data, color space is divides by discriminant analysis.

Fig. 3 Training data of components

ysis. Pixels group of nuclei, interstitium, cytoplasm, and
background are dened as Pn , Pi , Pc , and Pb . Color elements of a pixel p and the average of Px ({x|n, i, c, b})
Px dened as follow:
(
)
s(p)
p=
v(p)
( ) ( 1
)
sx
pPx s(p)
n

= 1
Px =
vx
pPx v(p)
nx

3. RESULT

Variance co-variance matrix Sx of group x is dened as

follow:
1
Sx =
{(p Px )(p Px )t }
nx

3.1 Background extraction

The histograms of some images background pixel
color Fig. 4.

pPx

Then, mahalanobis distance Dx (p) between a pixel p and

Px is dened as follow:

Dx (p) = (p Px )S 1 (p Px )

[

[

[







IUHTXHQF\




IUHTXHQF\

(b) Cytoplasm

IUHTXHQF\

(a) Interstitium

2.2.2 Nuclei extraction by Otsu method with Laplacian

histogram
Nuclei are stained darker color than other area. But,
simple Otsu method cant decide threshold when there
is too much difference between areas size in target
image. Therefore, Otsu method with Laplacian histogram method is used in this study. Laplacian histogram method utilizes edge-neighbors having pixels of
two groups equally. Extracting pixels having large edge
intensity, Laplacian histogram is made by these pixels.
Applying Otsu method on the histogram, threshold to divide nuclei and others is decided.









































The relation between pixel and groups is shown as follow:

(a) Red

Dx (p) Dn (p), Di (p), Dc (p), Db (p) p Px

(b) Green

(c) Blue

Fig. 4 Histogram of background area color

2.2 Dividing components by unsupervised learning

method
In this study, components are dividing on features of
colors and textures. First, background area is extracted
by xed color ranges. Second, nuclei, that have characteristic color and little area, are extracted by Otsu method
using Laplacian histogram. Third, cytoplasm and interstitium are tried to divide regions using texture features.

On Fig. 4, median colors intensity seems (R, G, B) =

(211.5, 208.5, 208.5). So color space of background is
set in 206 < R < 217, 203 < G < 214, 203 < B <
214. A extracted background image is shown on Fig. 5.
3.2 Nuclei extraction
Histograms of green intensity are shown on Fig. 6.
Extracted nuclei image is shown on g. 7.

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(a) Background

(b) Glandulars inner

(a) Nuclei mask

Fig. 7 Extracted nuclei image

Fig. 5 Extracted background and glandulars inner image

[

IUHTXHQF\

IUHTXHQF\




[

(a) Threshold = 1











(a) From original image



(b) Nuclei area














(b) Threshold = 5

(c) Result of
k-means

Fig. 8 2-D histogram of low edge intensity area



(b) From high edge-intensity

area

images have high-resolution: 0.275m/pixel. But they

were compressed and adjusted, so they have block noises.
Zoomed image and its Laplacian-ltered image is shown
on Fig. 10.
Blocks edges are shown Fig. 10. Then there are cyclical high edge-intensity areas in any component. In this
reason, reducing effect of block noises by ltering out
Fourier transform or wavelet transform is needed.

Fig. 6 Histogram of green intensity

3.3 Interstitium and cytoplasm extraction by textures
2-D histogram of saturation and value is shown on Fig.
8 when threshold of edge intensity image E is set on 1 and
5.
When Th was set on 1, the histogram has interstitium
and cytoplasm half-and-half. When Th is set on 5, cytoplasm is too much. So we applied k-means clustering
to the histogram on th=1. The result is shown on Fig.
9. Next, LUT by discriminant analysis from the cluster
is shown on Fig. 9 (a). And by comparison, LUT from
components extracted manually is shown on Fig. 9 (b).
Comparing Fig. 9 (a) and (b), result of k-means clustering couldnt divide color space of cytoplasm and interstitium well.

4.2 Problem of clustering

In this study, simple k-means clustering is used as unsupervised learning method. Simple k-means method is
usable when cluster number is known, but applying this
method was difcult when groups are near like our study.
So using different distance not Euclid or Manhattan distance but Mahalanobis genelralised distance is needed
for reduce effect of distribution.

5. CONCLUSION

4. DISCUSSION

Our study tried to divide components on biopsy images. Our enable to extract area of background and nuclei by xed color space or Otsu method with Laplacian
histogram. But on cytoplasm and interstitium, despite to
trying to extract by unsupervised learning with k-means
method and discriminant analysis, extracting accuracy
was not enough. Method to consider block noises and
color sample variance is needed.

In this instance, dividing accuracy between cytoplasm

and interstitium isnt enough. The reasons are considered.
4.1 Problem of image compressing
In this study, biopsy images were taken by digital slide
system at the Cancer Institute Hospital of JFCR. These

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(a) By k-means cluster

(b) By manual extracting area

Fig. 9 LUT of saturation and value

(a) Zoomed

(b) Laplacial (Red)

(c) Laplacial (Green)

(d) Laplacial (Blue)

Fig. 10 Zoomed image and its Laplacian-ltered image

REFERENCES
[1] T. Tanaka, A. Suzuki, Discriminant System for
Severity of Prostate Tumor, SICE society of pattern
measurement 78th, pp. 1-6, 2009.
[2] J. ZHANG, J. HU, Renal Biopsy Image Segmentation Based on 2-D Otsu Method with Histogram
Analysis, MEDICAL IMAGING TECHNOLOGY,
Vol. 27, pp. 185-192, 2009

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