Mueller Decomposition Images For Cervical Tissue: Potential For Discriminating Normal and Dysplastic States

Mueller decomposition images for cervical
tissue: Potential for discriminating normal and

dysplastic states
Prashant Shukla and Asima Pradhan
*

Department of Physics and Centre for Laser Technology, Indian Institute of Technology, Kanpur-208016
asima@iitk.ac.in
Abstract: We report the potential of Mueller decomposition images to
discriminate the normal against the dysplastic (precancerous) states in
cervical tissue. It is observed that in the epithelium region, depolarization
power is sensitive to morphological changes during progression from
normal to dysplastic state while retardance and diattenuation do not show
any significant change. These morphological changes have been correlated
with the microscopic images of the tissues. In contrast, it is the retardance
which reveals the morphological changes around the stromal region.
Additionally, we have evaluated the arithmetic mean of depolarization
power and retardance from their respective decomposed images and have
shown that this parameter has a potential to discriminate normal tissues
against dysplastic ones.
2009 Optical Society of America
OCIS codes: (110.0113) Imaging through turbid media; (170.6930) Tissue; (290.0290)
scattering; (290.5855) Scattering polarization; (290.7050) Turbid media

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#104234 - $15.00 USD Received 21 Nov 2008; revised 16 Jan 2009; accepted 22 Jan 2009; published 27 Jan 2009
(C) 2009 OSA 2 February 2009 / Vol. 17, No. 3 / OPTICS EXPRESS 1600
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1. Introduction
Precancerous stages (dysplasia) in cervical tissue are generally estimated by noting changes in
the epithelium region. The grade of dysplasia is decided by the thickness up to which
structural changes occur in the epithelium region [1]. However, it has been reported in the
literature that there is a distortion in the collagen fibers during the development of dysplastic
stage in stroma [2]. Such changes that occur in the stroma are generally not used for
histopathological analysis. Light scattering is known to reveal subtle structural changes in
cells [3]. Polarized light scattering in the form of Mueller matrix describes completely the
optical properties of any scattering medium like tissue, polystyrene micro-spheres etc.. In the
recent past, substantial theoretical [4-10] and experimental [11-17] studies based on the
Mueller matrix have been done. The 16 elements of Mueller matrix generated by recording
images of any scattering medium for various combinations of polarizer and analyzer can
provide information such as the size and refractive index of the scattering medium [16]. In
addition to these Mueller matrix elements, diattenuation, retardance and depolarization power
can also reveal more information about the structure and morphology of highly scattering
media such as biological tissues. These parameters (diattenuation, retardance and
depolarization power) for any scattering medium can be extracted by polar decomposition of
Mueller matrix given by Lu et. al. [4]. Liu et. al. have used polar decomposition algorithm for
rat skin and melanoma phantoms [14]. They have found that considerable information on the
morphological structures of any scattering medium can be extracted from Mueller
decomposition images in comparison to standard imaging techniques. Using 3x3 Mueller
decomposition technique, Swami et. al. have reported high retardance value shown by
collagen fibers extracted from eggshell membrane [17]. Recently Anastasiadou et. al. have
used DOP polarimetric technique for detection of cervical cancer and compared their results
with classical colposcopy. They concluded that DOP technique shows changes from normal to
cancer states and can be used for diagnosis purposes [18]. In a complex turbid medium like
biological tissue, many polarization effects occur simultaneously (the most common
polarimetry effects are depolarization, linear birefringence and optical activity). Thus the
Stokes parameter-based measure of degree of polarization represents the value of degree of
polarization resulting from several lumped, polarization effects. In contrast, the Mueller
matrix decomposition approach enables one to extract, quantify and interpret the individual
intrinsic polarimetry characteristics of tissue. Each of these, individual polarization
parameters, holds promise as a useful biological metric, as is also apparent from the results of
our studies. In the present work the polar decomposition algorithm on Mueller matrix [4] has
been used to study its potential to discriminate the normal against the precancerous stage
(dysplastic state) of human cervical tissues. Significant differences are observed in the
decomposition images of normal and dysplastic tissues of the same patient. Changes are
observed in the stroma of the cervical tissue through the retardance parameter while epithelial
changes are noticed through depolarization power.
2. Theory
2.1 Polar decomposition of the Mueller Matrices
Let M be a 4x4 Mueller matrix as given below

00 01 02 03
10 11 12 13
20 21 22 23
30 31 32 33
m m m m
m m m m
M
m m m m
m m m m
| | | | | | | |
| | | |
| | | |
= == =
| | | |
| | | |
| | | |
\ \ \ \
(1)

This Mueller matrix M can be decomposed into three elementary matrices representing a
depolarizer (M
), a retarder (M
R
) and a diattenuator (M
D
) [4]. The decomposition of Mueller
matrix depends upon the order in which the diattenuator, depolarizer and retarder matrices are
multiplied. Based on the order of these matrices, six possible decompositions can be
performed. Among these, the process in which the diattenuator matrix comes ahead of the
retardance and the depolarization matrix [M=M
M
R
M
D
] always leads to a physically
realizable Mueller matrix [5]. The decomposition process discussed in this paper is based on
this approach. Therefore, any Mueller matrix M can be mathematically decomposed as

R D
M=M M M (2)
where the depolarizing matrix M
accounts for the depolarizing effects of the medium, the

retarder matrix M
R
describes the effects of linear birefringence and optical activity, and the
diattenuator matrix M
D
includes the effects of linear and circular dichroism.
We can calculate directly diattenuation D from the Mueller matrix M as

( (( ( ) )) )
1 2
2 2 2
01 02 03
00
m +m +m
D=
m
(3)
From the Mueller matrix M, first we can construct a diattenuator Mueller matrix by taking
diattenuation vector D
as

01
02
00
03
m
1
D= m
m
m
| | | | | | | |
| | | |
| | | |
| | | |
\ \ \ \
(4)
Thus the first row of M gives the diattenuation vector. Then from this diattenuation vector the
diattenuator Mueller matrix can be constructed as

T
D
D
1 D
M =
D m
| | | | | | | |
| | | |
| | | |
\ \ \ \
(5)
Here
D
m is defined as

2 2 T
D

m = 1-D I +(1- 1-D ) DD (6)

where I is 3 3 identity matrix and
D
D =
D
| | | | | | | |
| | | |
| | | |
\ \ \ \
denotes the unit vector along D
.
Further a Mueller matrix M is defined based on M, as

-1
D
M = M M (7)
This M contains only retardance and depolarization and no diattenuation. M can be further
decomposed as a retarder followed by a depolarizer

R
R
|
R
1 0 1 0
M M =
P m 0 m
1 0 1 0
= =
P m m P m
= M
| | | | | | | | | | | | | | | |
| | | | | | | |
| | | | | | | |
\ \ \ \ \ \ \ \
| | | | | | | | | | | | | | | |
| | | | | | | |
| | | | | | | |
\ \ \ \ \ \ \ \

(8)

Here m is a 3x3 sub-matrix of M .

Equations (7) and (8) lead to

2
P- mD
P =
1- D

(9)

R
m m m = == = (10)
where polarizance vector P
can be expressed in terms of Mueller matrix elements as follows

10
20
00
30
m
1
P= m
m
m
| | | | | | | |
| | | |
| | | |
| | | |
\ \ \ \
(11)

Let
1
,
2
and
3
be the eigen values of
T
m (m) . m

has eigen values
1
,
2
and
3
.
m

can be obtained by

1
T 1
2
1 2 2 3 3 1
1 1 1 1
T
2 2 2 2
1 2 3 1 2 3
m [ m (m ) ( ) I ]
[{( ) ( ) ( ) }m (m ) ( ) I ]

= + + + = + + + = + + + = + + +
+ + + + + + + + + + + + (12)
If determinant of m is negative then minus sign is applied. So M

can be determined by Eqs.
(9) and (12). Once M

is determined then we can evaluate depolarization power as

tr(M )-1
= 1 -
3
(13)

Now M
R
can be obtained by

-1
R
M = M M (14)

From the retardance Mueller matrix, the retardance can be obtained by

-1 R
tr(M )
R=cos -1
2
( ( ( (
( ( ( (

(15)
Finally we obtain three parameters as:

1 2
2 2 2
01 02 03
00
1
D= m +m +m
m
(

(16)

( )
tr M -1
=1-
3
(17)

( )
R 1
tr M
R cos 1
2
(
=
(
(

(18)

A program was written in MATLAB (Math work) to decompose the Mueller matrices in
terms of the above parameters for each pixel of the illuminated region of CCD. Microscopic
images of these slides were also recorded to see the morphological changes histopathology. A
black mark was put on the tissue slide before recording the microscopic images so as to
ensure the illumination of the same spot for optical studies.
3. Experimental methods and materials
The samples used in this study were pathologically characterized, stained vertical sections of
human cervical tissues (containing both epithelial and stromal regions) for normal and
dysplastic states on glass slides. The lateral dimensions of tissue sample were 4 mm x 6 mm,
having thickness of 5 m. These samples were illuminated with He-Ne laser (Melles Griot,
20mW, = 632.8 nm) having spot size of 1.1 mm. Mueller images of these tissue slides were
recorded in the transmission mode on a CCD (Apogee 1E, USA) having resolution of 768 x
512 pixels. The incident beam was polarized using a linear polarizer (Glan Thompson having
extinction coefficient 100000:1) placed between the sample and the source. To generate
circularly polarized light, a quarter wave plate (05RP24-02, Newport) was introduced between
the polarizer and the sample. The transmitted light from the sample was collected by a convex
lens and after passing through subsequent polarization optics, was made to fall on the CCD.
Altogether 49 images were recorded for each sample using various combinations of polarizer,
analyzer and quarter wave plate to generate 16 elements of the Mueller matrix. Experiment
has been done on 10 pairs (normal and dysplasia) of cervical tissues. The experiment was
repeated three times for each sample to ensure reproducibility of experimental results.
4. Results and discussion
The experimental set-up was first calibrated by recording Mueller matrices for known optical
elements such as linear polarizer and air. The Mueller matrices for horizontal polarizer and air
are given below:
(a) For horizontal polarizer

1.0000 0.9791 0.0019 -0.0095
0.9682 0.9762 0.0170 -0.0308
-0.0475 -0.0355 -0.0079 -0.0181
0.0306 0.0197 -0.0144 -0.0162
| | | | | | | |
| | | |
| | | |
| | | |
| | | |
\ \ \ \

(b) For air

1.0000 0.0194 0.0351 0.0231
0.0201 0.9732 0.0191 0.0236
0.0040 0.0231 0.9632 0.0323
0.0087 0.0065 0.0265 0.9701
| | | | | | | |
| | | |

| | | |
| | | |

| | | |

\ \ \ \

Typical value of error in each element was found to lie between 1- 4%. After obtaining
satisfactory results from these standard measurements, the set-up was used to record Mueller
matrices for the samples investigated in this study. Figures 1 (a) and 1(b) show a typical
Mueller image (M
00
) and the corresponding microscopic image for the dysplastic state of the
cervical tissue respectively. The black point mark is visible in both images.

(a)
(b)
Fig. 1. (a) Mueller image M00 and (b) the microscopic image of a dysplastic
cervical tissue for the entire illuminated region.
(a) (b) (c)
Fig. 2. Images of (a) depolarization power (b) diattenuation (c)
retardance of a dysplastic cervical tissue section for the entire illuminated
region.

20 40 60 80 100120
20
40
60
80
100
120
0.4
0.5
0.6
0.7
0.8
0.9
1

20 40 60 80 100120
20
40
60
80
100
120
0
0.05
0.1
0.15
0.2

20 40 60 80 100120
20
40
60
80
100
120
0.5
1
1.5
2
2.5
3
Figure 2 represents the Mueller decomposition image in terms of depolarization power,
diattenuation and retardance for dysplastic cervical tissue section. It may be observed that the
black mark is reflected well in the depolarization power and the diattenuation images as
compared to the retardance image. This is expected since depolarization power and
diattenuation both reflect scattering and absorbance changes whereas retardance reflects the
birefringence and hence the black mark should not show up in the retardance image.
To correlate the alteration in morphology precisely at different regions of the normal and
dysplastic tissues, Mueller decomposition images were investigated separately for the
epithelium and stromal sides of tissue. Figure 3 shows the 2-D Mueller decomposition images
in terms of depolarization power for the epithelium region. The top of the image corresponds
to the outer region of the epithelium while the bottom corresponds to its lowest side (i.e. basal
layer which can be seen in the microscopic image also).

In the depolarization power images significant changes were observed in the epithelium
region of the dysplasia tissue as compared to the normal one. The value of depolarization
power in this region is large over the entire range of pixels along the vertical direction. The
large value of depolarization power in the dysplastic tissue indicates an increase in the value
of scattering coefficient. The increased value of scattering coefficient is equivalent to reduced
scattering mean free path which implies a higher density of scatterers. This is confirmed in the
microscopic image of the dysplastic tissue in Fig. (4), which shows the growth of cell density
in the epithelium region starting from the basal layer. A layered optical contrast is noticed in
the dysplastic state in contrast to an almost uniform density in the normal state. This variation
in the color contrast reflects the variation in the growth of cell density from basal to the outer
region of epithelium, during progression of dysplasia. It is observed in all the samples
consistently and may be a useful parameter for grading the various dysplastic states. The
number of samples studied here are not enough to confirm this. However, this gradation in
density is a definite parameter to discriminate dysplasia in general, from normal state with
100% sensitivity in 10 samples.

Pixel number
P
i
x
e
l

n
u
m
b
e
r

20 40 60 80
5
10
15
20
25
0.2
0.3
0.4
0.5
0.6
0.7
Pixel number
P
i
x
e
l

n
u
m
b
e
r

20 40 60 80
5
10
15
20
25
0.2
0.3
0.4
0.5
0.6
0.7
(a) (b)
Fig. 3. Typical depolarization power images of (a) normal and (b) dysplastic state in
the epithelium region of the cervical tissue section.

On the other hand, diattenuation and retardance images (not shown) were found to be
inconsistent to variations in normal and dysplastic tissues for the epithelium region. It thus
appears that morphological and structural changes of the epithelium region are better reflected
in terms of depolarization as compared to diattenuation and retardance in the Mueller
decomposition images.

In contrast, it is the retardance image of the stromal region in Fig. (5), which shows
consistent changes from the normal to the dysplastic state. The value of retardance is found to
be lower for the dysplastic state as compared to the normal counterpart. This reduction in the
value of retardance implies a decrease in the birefringence in the stromal region.
Birefringence arises due to fibrous structure of collagen present in the stromal region of
cervical tissue. Decrease in birefringence indicates deformation of the regular molecular
binding structure and even damage of the helix type molecules of collagen as reported in the
literature [2]. The diattenuation and depolarization images, however, do not show any
consistent changes in the stromal region. Therefore, morphological and structural changes of
the stromal region are less pronounced in terms of depolarization and diattenuation as
compared to retardance in the Mueller decomposition images.
The arithmetic mean of retardance, diattenuation and depolarization power have also been
calculated from their respective decomposed images to see whether the changes in these mean
values can discriminate among normal and dysplastic tissues. This arithmetic mean has been
Basal
layer
Basal
layer

(a) (b)
Fig. 4. Microscopic images of epithelial layer of (a) normal and (b) dysplastic
state.
Fig. 5. Typical retardance images of (a) normal and (b) dysplastic state in the
stromal region of the cervical tissue section.
(a) (b)
Pixel number
P
i
x
e
l

n
u
m
b
e
r

20 40 60 80
10
20
30
40
50
60
70
80
0.04
0.06
0.08
0.1
0.12
0.14
0.16
Pixel number
P
i
x
e
l

n
u
m
b
e
r

20 40 60 80
10
20
30
40
50
60
70
80
0.04
0.06
0.08
0.1
0.12
0.14
0.16
calculated by summing over the values of retardance, diattenuation and depolarization power
of each and every pixel of their corresponding decomposed images and then dividing the sum
by the total number of pixels. The fact that the retardance in the stromal region decreases from
normal to dysplastic tissues while the reverse happens in case of depolarization power in the
epithelium region, has been confirmed by this additional parameter.
Figures 6(a) and 6(b) show the mean values of depolarization power of epithelium region
and that of retardance of stromal region respectively. It is clear from the figure that the mean
values of depolarization power increase from normal to dysplastic tissues. It is found that the
mean value of depolarization power for normal tissues is generally less than 0.3 while for
dysplastic tissues, it is greater than 0.3 from the ten samples studied. Therefore, one can
observe that there is a clear demarcation for the mean values of depolarization power between
normal and dysplastic tissues which can serve as a benchmark for differentiating normal
against dysplastic tissues. However, for the 10 samples studied, such a distinct demarcation is
not apparent in the mean values of retardance of the stromal region as seen in Fig. 6(b).
Nevertheless, we observe that its value decreases from normal to dysplastic tissues
significantly for each patient, which is consistent with what we observed with the values of

retardance for stromal region in its corresponding decomposed images shown in Fig. 6(b). We
have found that the mean values of diattenuation did not show any consistent changes from
normal to dysplastic tissue, which is again consistent with what we observed in its
corresponding decomposed images.
5. Conclusion
In conclusion, our study shows that Mueller decomposition images have a potential to reveal
the structural and morphological changes in the dysplasia state of human cervical tissue
efficiently. We observe that in the epithelium region, depolarization power is sensitive to
morphological changes during progression from normal to dysplastic state while diattenuation
and retardance do not show any significant change. However, it is the retardance which
reveals the morphological changes around the stromal region. Changes in epithelium region
are conventionally used for diagnosis of dysplasia while changes in the stromal section are
generally not mentioned in the normal histopathology. However, with our observation of
stromal changes through retardance images, one can strengthen the diagnostic technique.
Additionally, we have defined a parameter in terms of mean values of depolarization power,
diattenuation and retardance calculated from their respective decomposed images. We have
shown that the mean values of depolarization power in epithelium region and retardance in
stromal region show significant changes from normal to dysplastic tissue while diattenuation
Fig. 6. (a) Mean value of depolarization power for the epithelium region and
(b) mean value of retardance for the stromal region.
(a)
(b)
0 1 2 3 4 5 6 7 8 9 10 11
0.24
0.28
0.32
0.36
0.40
0.44
0.48
0.52
Normal
Dysplastic
M
e
a
n

v
a
l
u
e

o
f

d
e
p
o
l
a
r
i
z
a
t
i
o
n

p
o
w
e
r
Patient number
0 1 2 3 4 5 6 7 8 9 10 11
0.05
0.06
0.07
0.08
0.09
0.10
0.11
Normal
Dysplastic
M
e
a
n

v
a
l
u
e

o
f

R
e
t
a
r
d
a
n
c
e
Patient number
does not show a consistent change. A consistent variation in the density, noticed in the
epithelial layer of dysplastic tissues may also be a useful parameter to determine various
stages of dysplasia. Further study on a larger number of samples is currently being pursued to
confirm this. We are of the opinion that this parameter which appears to show a cut-off in the
depolarization power values can be taken as a benchmark for discriminating normal tissues
against dysplastic one after performing a careful statistical analysis on a large number of
samples.
Acknowledgments
The authors would like to acknowledge Dr. Asha Agrawal for providing the tissue slides and
for fruitful discussions.

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Mueller decomposition images for cervical

tissue: Potential for discriminating normal and

accounts for the depolarizing effects of the medium, the

denotes the unit vector along D

can be expressed in terms of Mueller matrix elements as follows

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