You are on page 1of 6

Biospeckle: fractional Fourier transform point of view

William Lasso
a
, Eduardo A. Prez Caldern
b
, Nelali P. Castillo Serrano
b
, Leonardo Daz M.
a
, and
Cesar O. Torres*
a

a
Optics and Computer Science Laboratory, Popular University of Cesar, Valledupar, Colombia
b
University of Zulia, Maracaibo, Venezuela

ABSTRACT
The dynamic speckle analysis has been done because speckle interference began to be evaluated as an important source
of information, especially those related to biological samples under laser beam. The biospeckle is a phenomenon
produced naturally when biological material is illuminated by laser light. If it is possible to record the temporal history of
the phenomenon, it will be possible to identify, in a seed, areas with different activity. Biospeckle or dynamic speckle
can be used as a method for analysing activity, biologic or not, from materials illuminated with laser beam. The Spatial
Temporal Speckle (STS) contains data of time information of dynamic speckle and it is used as input for many
techniques allowing the analysis of the activity which is being monitored. This paper presents the results of analysing
biospeckle data using fractional order Fourier transform, coefficient filtering and reconstruction, with a novel approach
to increase the information obtained.

Keywords: Coherent light scattering, biological scattering centers, contrast, speckle size.
1. INTRODUCTION
Time history speckle pattern (THSP) has been used to characterize the activity in specimens, and the first account
registering its usage can be attributed to David Briers [1]. When coherent light crosses a medium having scattering
centers an ununiformly illuminated image is obtained, currently named speckled image, having a statistical distribution
of the intensity over the interference field. The speckled image appears as a result of the interference of the wavelets
scattered by the scattering centers, each wavelet having a different phase and amplitude in each location of the
interference field. An optical technique with potential use in biological metrology is the biospeckle; this effect (Braga Jr
et al., 2003) is a scattering phenomenon occurring when coherent light illuminates an active surface. Scattered waves are
Doppler shifted by movements of the scattering centres or other dynamic changes, such as refractive index time
variations, and beat in the detector thus producing low-frequency intensity variations. The surface appears to be covered
by tiny bright dots that fluctuate in a seemingly random way as for a boiling liquid. It has been used before for several
measurement applications in biology, medicine and industry.
The temporal variations of biospeckle are studied using some techniques as Inertia Moment [1], Wavelets based Entropy
[2] and Cross-Spectrum analysis [3]. All these methods have as input data the Spatial Temporal Speckle (STS) [4],
which is an image that represents the variation, along the time, of a region from the biospeckle of the material
illuminated with laser beam. In this work, we are going to consider the use of fractional order Fourier transform based
entropy as a measure of the activity following two approaches.
2. PRELIMINARIES
The application of dynamic laser speckle, or biospeckle laser BSL, in many areas of knowledge created naturally new
demands of research and developments [14], but always carrying out the results with the separation of the graphical to
the numerical approaches. The biospeckle samples can be obtained using different approaches. When only a small region
of the active sample is illuminated, one convenient way to register and store dynamic speckle information is through the
time history of the speckle pattern (THSP) suggested by Oulamara et al. [5]. Free propagation of the scattered light from
the sample is registered by a lens less CCD camera or a strongly defocused one. One column of the obtained image is
selected and consecutive ones are stored side by side in a composite image, the THSP.


*Email: cesartorres@unicesar.edu.co
8th Iberoamerican Optics Meeting and 11th Latin American Meeting on Optics, Lasers, and Applications,
edited by Manuel Filipe P. C. Martins Costa, Proc. of SPIE Vol. 8785, 8785G1
2013 SPIE CCC code: 0277-786X/13/$18 doi: 10.1117/12.2026264
Proc. of SPIE Vol. 8785 8785G1-1
Downloaded From: http://proceedings.spiedigitallibrary.org/ on 12/12/2013 Terms of Use: http://spiedl.org/terms



Every pixel in the CCD receives, but for negligible variations in the sensitivity vector, different instantiations of
essentially the same phenomenon. So, many samples are simultaneously registered and an efficient use of statistics can
be accomplished. The incoming images are digitized by a frame grabber and stored and processed by a personal
computer. Care is taken to maintain the laser intensity constant during data acquisition.

The Fresnel and Fraunhofer diffraction equations can be derived from a transmission aperture diffraction model
(TADM), where the observation plane parallels the aperture plane; as a consequence, if the observation plane is not
parallel to the aperture plane, or, for a reflective surface diffraction model (RSDM) the observation plane is not parallel
to the object plane, the Fresnel and Fraunhofer diffraction equations become incapable of evaluating the complex-
amplitude distribution at the observation plane. This can be attributed to two considerations: (1) the coordinate system
rotation between the observation plane and the object plane, (2) variations of the near-field approximation caused by the
coordinate system rotation. There is also a distinct difference in the calculation of the inclination factor using the RSDM
and the TADM. When a weakly scattering object with a reflective surface is illuminated by a coherent oblique plane
wave, choosing an observation plane perpendicular to the direction of the reflected light by the object plane, a partially
developed speckle pattern will occur in this plane. To evaluate the speckle field at the observation plane, a new
diffraction equation need to be established. Yet to our best knowledge, few studies have been focused on this point.
Cheng [5] proposed an interesting RSDM by approximating the actual reflective surface with a series of step-like planes
and gave a discrete expression of the diffraction field under the Fraunhofer approximation. Allardyce et al. [8] analyzed
the diffraction from a rough reflective surface, using geometrical optics to acquire the emergent light field of the surface
and employing the Fraunhofer diffraction equation. New theoretical results of the diffraction of a rough surface under
the case in which the observation and the object planes are parallel to each other. Yamaguchi et al. [8] presented a
diffraction equation of a reflective surface, taking only the coordinate system rotation into account.
2.1 Analysing the biological activity by speckle properties
Surface roughness measurements of metallic components are of great importance in the mechanical machining field.
Since 1972 when Sprague [1] applied white speckle techniques to surface roughness measurements, many investigators
have investigated this subject [2-8]. Speckle patterns contain information about the microscopic structures of a rough
surface. In order to study the relationships between the statistical properties of speckle patterns with surface roughness,
the complex-amplitude distribution of the speckle field should be obtained. To achieve this, the corresponding diffraction
equation describing the propagation of diffracted wave from the rough surface to the observation plane should be
constructed. In former studies, the Fresnel and Fraunhofer diffraction equations were used to calculate the speckle field
[6-8] with the consideration that the observation plane is in parallel with the object plane in their experimental setups.
The purpose of this paper is to derive corresponding diffraction equations to describe the diffraction from a weak
scattering reflective surface under a coherent plane wave incidence. We consider the difference between the reflective
surface diffraction model (RSDM) and the transmission aperture diffraction model (TADM) and reached a general
diffraction equation in terms of the fractional Fourier transform and mathematical modeling of dynamic speckle.
2.2 General diffraction equation for a inclined surface
The integral of Fresnel-Kirchhoff diffraction can be written as:
( ) ( )
( )
ds
r
ikr
P U P U
01
01
1
'
0
exp

=
(1)
Where is the surface of the integral, and is chosen to be modeled mathematically as an inclined aperture; this opening
can be written as:
( ) ( ) ( )


=
2
, cos , cos exp 1
) ( '
21 01
21
21
1
r n r n
r
ikr A
i
P U
r r r r

, consequently the Equation 1 can thus be written as:


( )
( ) ( ) ( )
ds
r n r n
r
ikr
P U
i
P U

2
, cos , cos exp 1
) (
21 01
01
01
1 0
r r r r

(2)
Proc. of SPIE Vol. 8785 8785G1-2
Downloaded From: http://proceedings.spiedigitallibrary.org/ on 12/12/2013 Terms of Use: http://spiedl.org/terms


Where the expression ) (
1
P U is the wave field in the diffraction point and ) (
0
P U is the wave field in the observation
point
0
P , and the parenthetical term expression (2) represents the diffraction angle and the slope factor for the model
respectively. Considering in the point
1
P the corresponding expression for
01
r
r
and the element of area ds in the equation
(2) and also substituting ( )
1
P U , and ( )
01
, cos r n
r r
the diffraction general equation is obtained for an inclined surface:
( ) ( )
( ) ( ) ( )
( ) ( ) ( )
( ) ( ) ( )
( ) ( ) ( )
1 1
2
0 1
2
0 1
2
0 1
0 1 0 1 0 1
2
0 1
2
0 1
2
0 1
2
0 1
2
0 1
2
0 1
1 1 0 0
cos sin
exp
cos sin
2
exp
2
1
dy dx h
z z y y x x
z z y y h x x h
z z y y x x
z z y y x x ik
z x i A
i
P U
x
y x
D

+ +
+ +
+
+ +

+ +

(3)

Where D defines the region of integration in the median plane.
2.3 Near field diffraction
As seen from equation (3) is very wasteful calculate the diffraction of light that should be roughly taken into account in
practical utilities, however considering the geometric arrangement of Figure 1, and based on the paraxial approximation,
where it is assumed that the distance L is very large compared with the linear dimensions of the region of maximum
illumination and the observation point
0
P is very close to the z axis, if it is further necessary approximations of
01
r
r
and
the terms of the partial derivation of
x
h ,
y
h , and substituted in equation (3) gives an approximate expression for the
diffraction equation for a reflective inclined surface.

Fig. 1. Geometrical arrangement
( ) ( )
( )
( )
1 1
2
0
2
0
2
0
1 0
2
0
2
0
2
0
0 1 0 1
2
0
2
0
2
0
2
1
2
1
1 1
2
0
2
0
2
0
2
0
2
0
2
0
2
0
2
0
2
0
0 0
0 0 0 0
2
exp
2
exp
exp cos sin
2
exp
exp
cos
2
, ,
dy dx
z y x
z z
i
z y x
y y x x
i
z y x
y x
i z x i
z y x
z y x ik
z y x
z
i
A
z y x U P U
D

+ +

+ +
+

+ +
+


+ +
+ +

+
+ +
= =

(4)
Proc. of SPIE Vol. 8785 8785G1-3
Downloaded From: http://proceedings.spiedigitallibrary.org/ on 12/12/2013 Terms of Use: http://spiedl.org/terms


Making a coordinate conversion, cos sin
0
+ = L x , =
0
y , sen L z = cos
0
and entering the aperture
function ( ) y x P , the equation (4) yields equation near field diffraction for a rough surface inclined.
( )
( )
( ) ( )
( )
1 1
2 2 2
1 1
2 2 2
2
1
2
1
1
2 2 2
1 1 1
2 2 2
2 2 2
2 2 2
0
cos sin 2
exp exp
sin cos
cos
2
exp sin
2
exp ,
exp
cos
sin cos
2
,
dy dx
L
y L x
i
L
y x
i
z
L
L
i x i y x P
L
L ik
L
L
i
A
U

+ +
+ +

+ +
+

+ +

+

+ +
+ +

+
+ +

=


(5)
Considering that:
( ) ( )

+ +

+ =
1
2 2 2
1 1 1 1 0
sin cos
cos
2
exp , , z
L
L
i y x P y x U

Equation (5) can be expressed in terms of a


fractional Fourier transform order, as shown in equation (6):
( )
( ) ( )
( ) ( )

+ +
+ +

+ +
+ +

+
+ +

=



sin
2
exp ,
cos sin
exp
exp
cos
sin cos
2
,
1 1 1 0
2 2 2
2 2
2 2 2
2 2 2
2 2 2
0
x i y x U
L
L
i
L
L ik
L
L
i
A
U
(6)
The expression (6) means that the aperture or pupil function is mounted on a phase of the form:
( )

sin
2
exp
1
x i
,
which corresponds to a spherical wave, Equation (6) establishes a relationship of fractional Fourier transform between
the complex amplitude distributions of field. Considering equation (6) approaches necessary to help describe the near-
field diffraction, one can obtain the standard form of the Fresnel diffraction equation for an inclined and reflective
surface, ie:
( )
( ) ( )
( ) ( )

+ +

+

+ +
+ +

+ +
+ +

+
+ +

=

sin
2
exp
sin cos
cos
2
exp ,
cos sin
exp
exp
cos
sin cos
2
,
1 1
2 2 2
1 1
2 2 2
2 2
2 2 2
2 2 2
2 2 2
0
x i z
L
L
i y x P
L
L
i
L
L ik
L
L
i
A
U
(7)
In the same way, considering the far-field diffraction ( )

L
y x +
max
2
1
2
1
allows to obtain the standard form of the
Fraunhofer diffraction equation for an inclined and reflective surface, ie:
( )
( )
( ) ( )

+ +

+
+ +
+ +

+
+ +

=

sin
2
exp
sin cos
cos
2
exp ,
exp
cos
sin cos
2
,
1 1
2 2 2
1 1
2 2 2
2 2 2
2 2 2
0
x i z
L
L
i y x P
L
L ik
L
L
i
A
U
(8)
In the particular situation where there is no inclination 0 = , the equations (7) and (8) yields:

( )
( )
( ) { }
1 1 0
2 2
0
, exp
exp
, y x U
L
i
L
ikL
i
A
U

+
=
(9)
( )
( )
( ) { }
1 1 0
0
,
exp
, y x U
L
ikL
i
A
U =

(10)
Proc. of SPIE Vol. 8785 8785G1-4
Downloaded From: http://proceedings.spiedigitallibrary.org/ on 12/12/2013 Terms of Use: http://spiedl.org/terms


Equations (9) and (10) are precisely classic expressions corresponding to Fresnel diffraction and Fraunhofer diffraction
plane and the observation plane is not inclined.
3. A BIOLOGICAL SURFACE ILLUMINATED BY A LASER BEAM: THEORY
A biological surface illuminated by a laser beam can be modeled as the product of two components:

a) ( )
1 1 1
, , z y x C , that describes the roughness of the sample which consists of elementary cells with random
distribution of size and orientation.
b) ( ) t y x D , ,
1 1
, that describes the instantaneous location of small and moving particles inside the cells.

The speckle pattern observed in the observation plane at time t result from the light diffracted by the instantaneous
distribution of scatterers. The three-dimentional diffusing target is defined by the product:
( ) ( ) ( ) t y x D z y x C t z y x P , , , , , , ,
1 1 1 1 1 1 1 1
= .

( ) ( ) ( )
1 1 1 1 1 1 1
, , , , z y C z x C z y x C
y x
= , denotes the stationary roughness function with respect to the average plane of the
sample. Assuming a rectangular aperture:

( ) ( )

<

= =
a x if
a x if
x rect z x C
x
1
1
1 1 1
0
1
, ( ) ( )

<

= =
b y if
b y if
y rect z y C
y
1
1
1 1 1
0
1
, (11)

And the aperture function can be expanded as the sum of complex Gaussian functions with finite numbers and given by

( )

= =

=
N
l
l
l
N
n
n
n
y
b
B
A x
a
B
A z y x C
1
2
1 2
1
2
1 2 1 1 1
exp exp , , (12)
( ) t y x D , ,
1 1
is the temporal distribution of the scatterers defined by their individual location in the ( ) y x, plane
assuming displacement dz to be small, them:

( ) ( ) ( )
i i
y y x x t y x D = , ,
1 1
(13)

At time ( ) t t + this function is:

( ) ( ) ( ) ( ) ( ) t v y y t v x x t t y x D
yi i xi i
+ + = + , ,
1 1
(14)

xi
v and
yi
v are the average speeds of the particle in the ( ) y x, plane. The scattered field amplitude becomes:

( )
( ) ( )
( ) ( ) ( ) ( ) ( ) ( )

+ +

+ + +

+ +
+ +

+ +
+ +

+
+ +

=

= =

sin
2
exp
sin cos
cos
2
exp , , exp exp
cos sin
exp
exp
cos
sin cos
2
,
1 1
2 2 2
1 1
1
2
1 2
1
2
1 2
2 2 2
2 2
2 2 2
2 2 2
2 2 2
0
x i z
L
L
i t y x D t v y y t v x x y
b
B
A x
a
B
A
L
L
i
L
L ik
L
L
i
A
U
yi i xi i
N
l
l
l
N
n
n
n
(15)



Proc. of SPIE Vol. 8785 8785G1-5
Downloaded From: http://proceedings.spiedigitallibrary.org/ on 12/12/2013 Terms of Use: http://spiedl.org/terms



CONCLUSIONS

Based on the integral of the Fresnel-Kirchhoff diffraction, we have obtained a diffraction basic integrated for a reflective
inclined surface, in terms of the fractional Fourier transform and mathematical modeling of dynamic speckle, where the
difference in the calculation of tilt factor between the model has been taken into account. As a fundamental result found
a general equation of diffraction for an inclined surface and reflective under illumination (typical speckle patterns) of a
plane wave coherent, considering the effects of rotation of the coordinate system on the near-field approximation and
introducing the consideration of small inclinations, the diffraction near field and far field are obtained and presented as
special cases of equation (9-10). Thus the resulting expression is more general than the classical equations of Fresnel
diffraction and Fraunhofer for typical speckle patterns, It should be noted that the light fields emerging microscopic
equations are contained in these diffraction, and refer to the height distribution of the surface considered


REFERENCES
[1] Sprague RA. Surface roughness measurement using white light speckle. Appl. Opt. 1972; 11(12):28116.
[2] Chandley PJ. Surface roughness measurements from coherent light scattering. Opt. Quantum Electron 1976;8:323
7.
[3] Pedersen HM. Object-roughness dependence of partially developed speckle patterns in coherent light. Opt Commun
1976;16(1):637
[4] Leonard LC, Toal V. Roughness measurement of metallic surfaces based on the laser speckle contrast method. Opt
Lasers Eng 1998;30:43340.
[5] Yamaguchi I, Kobayashi K, Yaroslavsky L. Measurement of surface roughness by speckle correlation. Opt Eng
2004;43(11):275361.
[6] Goodman JW. Dependence of image speckle contrast on surface roughness. Opt Commun 1975;14(3):3247.
[7] Ohtsubo J, Asakura T. Statistical properties of laser speckle produced in the diffraction field. Appl Opt
1977;16(6):174253.
[8] Zhao Gao, Xuezeng Zhao. Diffraction equations for weak scattering reflective surface. Optics & Laser Technology,
May 2008.

Proc. of SPIE Vol. 8785 8785G1-6
Downloaded From: http://proceedings.spiedigitallibrary.org/ on 12/12/2013 Terms of Use: http://spiedl.org/terms