CHAPTER 1

INTRODUCTION
Cancer is defined as the abnormal growth of tissues. Prostate cancer is a form of cancer that
develops in the prostate, a gland in the male reproductive system. Most prostate cancers are slow
growing. The cancer cells may spread from the prostate to other parts of the body; particularly,
the bones and lymph nodes. Rates of detection of Prostate cancer vary widely across the world.
The Prostate cancer is the second leading cause of cancer-related death among men and is the
most commonly diagnosed cancer in men.

Image segmentation is the division of an image into regions or categories, which correspond to
different objects or parts of objects. Every pixel in an image is allocated to one of a numberof
these categories. A good segmentation is typically one in which:- a.) pixels in the same category
have similar greyscale of multivariate values and form a connected region, b.) neighbouring
pixels which are in different categories have dissimilar values.

Segmentation is a fundamental operation on images that partitions the image into homogeneous
segments or regions to ease the analysis of images. It is the process of grouping pixelstogether,
with each group having at least one or more features in common. The features may bebrightness,
color, motion, texture etc. Boundary detection is an integral part of this processsince it helps to
identify the individual segments themselves. Segmentation is represented as aclustering problem
in statistics and can be used for 3D reconstruction of objects when appliedto a stack of images
such as in Medical Imaging.

1.1 Role of computer in medical imaging

In computer vision, segmentation is widely used to separate the foreground from the back ground
and for feature extraction, face detection, video compression etc. It is one of the oldestand most
extensively studied problems with Brice and Fennema contributing significantly by beginning to
analyze scenes using regions, in 1970. Early techniques of segmentation used region splitting or
merging while recent techniques often tend to optimize some global criterion.This report
describes the technique for detecting cancer cell using image segmentation in MATLAB. A new
algorithm is implemented to successfully segment intensity inhomogeneous images.
Segmentation of cancer cells images in prostate gland of male reproductive organusing this
proposed algorithm is shown and the results are discussed.

A medical image archiving system and method can be employed to understand image
segmentation. A medical image archiving system receives analog NTSC or PAL video from a
medical imaging device and converts it to a digital format for storage. The storage can be via
local hard disc drive, or CD Writer, or other optical storage medium, or via Local or Wide area
network storage to a remote electronic storage medium. The system includes an integral Web
server to permit easy access over a network using a browser. When an image is stored on a CD, it
can be stored as a session and the CD closed to prevent further Writing.

Fig1.1. Scheme of medical imaging archiving system.

1.2 Aim of the Project:

The aim of this project is to show how to detect a cell using image segmentation. An object can
be easily detected in a image if the object has sufficient contrast from the background. In this
project we detect prostate cancer cells using edge detection and basic morphology. These days,
the scope of medical imaging is increasing rapidly with the collaboration of engineering in this
area. Thus this project is a step forward in the field of medical imaging and letting identify the
future scope of the biomedical applications in the field of digital image processing.
CHAPTER 2

LITERATURE SURVEY

The aim of this section is to relate reader with the survey of basic processes and literature of
current research in the field of image sementation in medical imaging to detect a cancer cell.

2.1 Introduction:

For many years, research efforts have been focused on accurate methods of differentiating
indolent from more aggressive cancers. Since the original description by Gleasonof a pathologic
grading system, other methods of predicting cancer aggressiveness, using nuclear morphometry
as well as various molecular markers which include DNA ploidy, have been reported. These
methods have been based primarily on the analysis of fixed, histologic sections of prostate
cancer. By concentrating research efforts solely on the analysis of nonliving tissue, we are
ignoring certain aspects of prostate cancer that may be important in determining the metastatic
potential of individual tumors. Dynamic analysis of live cells to quantify various motility
characteristics has been instrumental in understanding ciliary function, cell development, early
physiologic cellular reactions, nuclear organization, cytoplasmic forces, neuronal death, and the
metastatic potential of animal models of prostate cancer. Common methods of imaging these
cells for motilityanalysis have been brightfield, fluorescence, phase contrast, modulation
contrast, and differential interference contrast microscopy. Not all of these imaging modalities
are optimal for live prostate cells. For instance, the transparency of cells often leads to low-
contrast images if bright field microscopy is used. Vital dyes have the potential for influencing
cell motility processes and decay of the signal occurs rapidly, thus limiting the extent to which
fluorescence microscopy can be employed to quantify cell motility. Phase contrast techniques
accentuate the boundaries of flat surfaces of prostate cells with a white halo, creating an
artifact that interferes with further analysis of cell shape. In consequence of these shortcomings,
modulation contrast (Hoffman) and differential interference contrast (DIC) microscopy are often
used to achieve high-contrast images of motile prostate cells. Hoffman microscopy detects
optical density gradients using a spatially varying transmittance filter (modulator).
This filter causes the modulation of opposite gradients above and below average background
intensity, which produces a three-dimensional appearance to a cell image. DIC microscopy also
gives a three-dimensional appearance to a cells image, but does so by using birefringenceoptics
consisting of Nomarski prisms and a polarizer. Live cell boundary detection, i.e., image
segmentation, is a central requirement of any quantitative motility analysis.
Special attributes of Hoffman and DIC images undermine the use of segmentation techniques
that have been developed for analysis of brightfield and fluorescence images. For example, the
intensity of cell borders in Hoffman images can be lighter or darker (or both) than the
surrounding background intensity (asymmetric intensity border), which makes segmentation
difficult. DIC images place an even greater demand on segmentation algorithms due to their
resolution of common delicate cell components (e.g., membrane ruffles, pseudopodal extensions,
undulations) in addition to the asymmetric intensity borders. As a result, effective use of
Hoffman and DIC images requires modification of many segmentation techniques, including
image thresholding, first- and second-order statistical analyses, B-splines, color feature analysis,
and fractal features. In order to segment the borders of cultured cells in Hoffman and DIC
images, we developed specialized algorithms that could be utilized in an automated system for
performing cell motility and morphometry analyses. Our focus has been on the use of segmented
cell information in the prediction of metastatic potential of ratprostatic adenocarcinoma cell
lines. Previously, Partin et al. (19) developed a Fourier analysis method that used a sequence of
spatial and temporal Fast Fourier Transforms to describe quantitatively the cells dynamic shape
change, based on temporal alterations in the boundary. This Fourier analysis scheme was applied
to in vitro cells from low and high metastatic Dunning sublines of the rat prostatic
adenocarcinoma (19). This study showed a strong correlation between the calculated Fourier
measurements and metastatic potential. Widespread use of this Fourier methodology has been
partially limited because earlier quantitative analyses required timeintensive and subjective
manual tracing of cell boundariesfrom microscopic images. To eliminate the tedious process of
manual cell tracing, we developed a segmentation algorithm for a cell motility analysis system.
This approach has the further benefit of providing an objective, reproducible method of
determining cell boundaries for our motility system and for other systems involving cell image
analysis. We examine the segmentation of cultured motile prostate cells, imaged using both
Hoffman and DIC video microscopy. Features are extracted from segmented cell boundaries and
used to characterize the morphometry of these motile cells, which can be utilized in prostate
cancer research. Features extracted from cell boundaries derived using our segmentation
algorithms are comparedwith those derived from manual cell tracings.

2.2 Images and Pictures:

Human beings are predominantly visual creatures. We not only look at things to identify and
classify them, but we can scan for differences, and obtain an overall rough feeling for a scene
with a quick glance.

Humans have evolved very precise visual skills: we can identify a face in an instant; we can
differentiate colours; we can process a large amount of visual information very quickly.

An image is a single picture which represents something. It may be a picture of a person, of

people or animals, or of an outdoor scene, or a microphotograph of an electronic component, or
the result of medical imaging.

2.2.1 Type of Images:

(i) Binary: Each pixel is just black or white. Since there are only two possible values for each
pixel, we only need one bit per pixel. Such images can therefore be very efficient in terms of
storage. Images for which a binary representation may be suitable include text (printed or
handwritten), fingerprints, or architectural plans.

(ii) Greyscale: Each pixel is a shade of grey, normally from 0 (black) to 255 (white). This range
means that each pixel can be represented by eight bits.

(ii) RGB (or True) Images: Here each pixel has a particular colour; that colour being described
by the amount of red, green and blue in it. If each of these components has a range0 to 255, this
gives a total of 2553 = 1, 67, 77,216 different possible colours in the image. This is enough
colours for any image. Since the total number of bits required for each pixel is 24, such images
are also called 24-bit colour images. Such an image may be considered as consisting of a stack of
three matrices; representing the red, green and blue values for each pixel. This means that for
every pixel there correspond three values.
 (iv) Indexed: Most colour images only have a small subset of the more than sixteen million
possible colours. For convenience of storage and file handling, the image has an associated
colour map, or colour palette, which is simply a list of all the colours used in that image. Each
pixel has value which does not give its colour (as for an RGB image), but an index to the colour
in the map. It is convenient if an image has 256 colours or less, for then the index values will
only require one byte each to store.

2.2.2 Image Processing:

Image processing is any form of signal processingfor which the input is an image, such as
a photograph or video frame; the output of image processing may be either an image or a set of
characteristics or parameters related to the image.

Digital image processing is the use of computer algorithms to perform image processing on
digital images. It allows a much wider range of algorithms to be applied to the input data and
can avoid problems such as the build-up of noise and signal distortion during processing. Since
images are defined over two dimensions (perhaps more) digital image processing may be
modeled in the form of multidimensional systems.

Types of Image Processing:

(i) Image enhancement: This refers to processing an image so that the result is more suitable for a
particular application. Examples include:
Sharpening or de-blurring an out of focus image.
Highlighting edges.
Improving image contrast or brightening an image.
Removing noise.
(ii) Image restoration: This may be considered as reversing the damage done to an image by a
known cause. For example:
Removing of blur caused by linear motion.
Removal of optical distortions.
Removing periodic interference.
 (iii) Image segmentation: This involves subdividing an image into constituent parts, or isolating
certain aspects of an image. For examples:
Finding lines, circles, or particular shapes in an image.
In an aerial photograph, identifying cars, trees, buildings, or roads.

2.2.3 Applications:

Image processing has an enormous range of applications; almost every area of science and
technology can make use of image processing methods. Here is a short list just to give some
indication of the range of image processing applications.

(i) Medicine:

Inspection and interpretation of images obtained from X-rays, MRI or CAT scans,
Analysis of cell images, of chromosome karyotypes.
(ii) Agriculture:

Satellite/aerial views of land, for example to determine how much land is being used for different
purposes, or to investigate the suitability of different regions for different crops,

Inspection of fruit and vegetables distinguishing good and fresh produce from old.
(iii) Industry:

Automatic inspection of items on a production line, Inspection of paper samples.

(i) Law enforcement:

Fingerprint analysis.
Sharpening or de-blurring of speed-camera images.

2.3 Various Techniques for cell detection:

2.3.1 METHODS:

Several common approaches have appeared in the recent literature on medical image
segmentation. We define each method, provide an overview of its implementation, and discuss its
advantages and disadvantages. Although each technique is described separately, multiple
techniques are often used in conjunction for solving different segmentation problems. We divide
segmentation methods into eight categories: (a) thresholding approaches, (b) region growing
approaches, (c) classifiers, (d) clustering approaches, (e) Markov random field (MRF) models,
(f) artificial neural networks, (g) deformable models, and (h) atlas-guided approaches.

Thresholding:
Thresholding approaches segment scalar images by creating a binary partitioning of the image
intensities. A thresholding procedure attempts to determine an intensity value, called the
threshold, which separates the desired classes. The segmentation is then achieved by grouping all
pixels with intensities greater than the threshold into one class and all other pixels into another
class.Determination of more than one threshold value is a process called multithresholding.
Thresholding is a simple yet often effective means for obtaining a segmentation of images in
which different structures have contrasting intensities or other quantifiable features. The partition
is usually generated interactively, although automated methods do exist. Thresholding is often
performed interactively, based on the operators visual assessment of the resulting segmentation.
Thresholding is often used as an initial step in a sequence of image-processing operations. It has
been applied in digital mammography, in which two classes of tissue are typically present
healthy and tumorous. Its main limitations are that, in its simplest form, only two classes are
generated, and it cannot be applied to multichannel images. In addition, thresholding typically
does not take into account the spatial characteristics of an image. This causes it to be sensitive to
noise and intensity inhomogeneities, which can occur in MR images.
Both of these artifacts essentially corrupt the histogram of the image, making separation more
difficult. For these reasons, variations on classical thresholding have been proposed for medical-
image segmentation that incorporate information based on local intensities and connectivity.

Region Growing:
Region growing is a technique for extracting an image region that is connected based on some
predefined criteria. These criteria can be based on intensity information and/or edges in the
image. In its simplest form, region growing requires a seed point that is manually selected by an
operator and extracts all pixels connected to the initial seed based on some predefined criteria.
 Figure 2.3: Feature space methods and region growing. (a) Histogram showing three
apparent classes. (b) 2-D feature space. (c) Example of region growing.
For example, one possible criterion might be to grow the region until an edge in the image is
met. This is depicted in Figure 2.3b, in which region growing has been used to isolate one of the
Structures.

Classifiers:
Classifier methods are pattern recognition techniques that seek to partition a feature space
derived from the image by using data with known labels. A feature space is the range space of
any function of the image, with the most common feature space being the image intensities
themselves. All pixels with their associated features on the left side of the partition would be
grouped into one class.
Classifiers are known as supervised methods because they require training data that are manually
segmented and then used as references for automatically segmenting new data. There are a
number of ways in which training data can be applied in classifier methods. A simple classifier is
the nearest-neighbor classifier, in which each pixel is classified in the same class as the training
datum with the closest intensity. The k-nearest-neighbor classifier is a generalization of this
approach, in which the pixel is classified into the same class as the majority of the k-closest
training data. The k-nearest-neighbor classifier is considered a nonparametric classifier because
it makes no underlying assumption about the statistical structure of the data. Another
nonparametric classifier is the Parzen window, in which the classification is made by a weighted
decision process within a predefined window of the feature space, centered at the unlabeled pixel
intensity.
A commonly used parametric classifier is the maximum-likelihood or Bayes classifier. It
assumes that the pixel intensities are independent samples from a mixture of probability
distributions, usually Gaussian. This mixture, called a finitemixture model, is given by the
probability density function:
yj
K
k
f(yj; , = k f k ;
k=1

For Gaussian mixtures, this means estimating K-means, covariances, and mixing coefficients.
Classification of new data is obtained by assigning each pixel to the class with the highest
posterior probability. When the data truly follow a finite Gaussian mixture distribution, the
maximum-likelihood classifier can perform well and is capable of providing a soft segmentation
composed of the posterior probabilities.
Standard classifiers require that the structures to be segmented possess distinct quantifiable
features. Because training data can be labeled, classifiers can transfer these labels to new data as
long as the feature space sufficiently distinguishes each label as well. Being noniterative,
classifiers are relatively computationally efficient, and, unlike thresholding methods, they can be
applied to multichannel images. A disadvantage of classifiers is that they generally do not
perform any spatial modeling. This weakness has been addressed in recent work extending
classifier methods to segmenting images that are corrupted by intensity inhomogeneities.
Another disadvantage is the requirement of manual interaction to obtain training data. Training
sets can be acquired for each image that requires segmenting, but this can be time consuming and
laborious. On the other hand, use of the same training set for a large number of scans can lead to
biased results that do not take into account anatomical and physiological variability between
different subjects.

Clustering:
Clustering algorithms essentially perform the same function as classifier methods without the use
of training data. Thus, they are termed unsupervised methods. To compensate for the lack of
training data, clustering methods iteratatively alternate between segmenting the image and
characterizing the properties of each class. In a sense, clustering methods train themselves, using
the available data. Three commonly used clustering algorithms are the K-means or ISODATA
algorithm, the fuzzy c-means algorithm, and the expectation-maximization (EM) algorithm. The
K-means clustering algorithm clusters data by iteratively computing a mean intensity for each
class and segmenting the image by classifying each pixel in the class with the closest mean. The
fuzzy c-means algorithm generalizes the K-means algorithm, allowing for soft segmentations
based on fuzzy set theory. The EM algorithm applies the same clustering principles with the
underlying assumption that the data follow a Gaussian mixture model. It iterates between
computing the posterior probabilities and computing maximum likelihood estimates of the
means, covariances, and mixing coefficients of the mixture model.
Although clustering algorithms do not require training data, they do require an initial
segmentation (or, equivalently, initial parameters). The EM algorithm has demonstrated greater
sensitivity to initialization than the K-means or fuzzy c-means algorithm. Like classifier
methods, clustering algorithms do not directly incorporate spatial modeling and can therefore be
sensitive to noise and intensity inhomogeneities. This lack of spatial modeling, however, can
provide significant advantages for fast computation. Work on improving the robustnessof
clustering algorithms to intensity inhomogeneities in MR images has demonstrated excellent
success. Robustness to noise can be incorporated byMRF modeling as described in the next
section.

Markov Random FieldModels:

MRF modeling itself is not a segmentation method but a statistical model that can be used within
segmentation methods. MRFs model spatial interactions between neighboring or nearby pixels.
These local correlations provide a mechanism for modeling a variety of image properties. In
medical imaging, they are typically used because most pixels belong to the same class as their
neighboring pixels. In physical terms, this implies that any anatomical structure that consists of
only one pixel has a very low probability of occurring under an MRF assumption. MRFs are
often incorporated into clustering segmentation algorithms such as the K-means algorithm under
a Bayesian prior model. The segmentation is then obtained by maximizing the a posteriori
probability of the segmentation, given the image data. This maximization can be achieved by
iterative methods such as iterated conditional modes or simulated annealing. The segmentation
does not exhibit as many small, disconnected regions as the non-MRF result. A difficulty
associated with MRF models is proper selection of the parameters controlling the strength of
spatial interactions. A setting that is too high can result in an excessively smooth segmentation
and a loss of important structural details. In addition, MRF methods usually require
computationally intensive algorithms.

Artificial Neural Networks:

Artificial neural networks (ANNs) are parallel networks of processing elements or nodes that
simulate biological learning. Each node in an ANN is capable of performing elementary
computations. Learning is achieved through the adaptation of weights assigned to the
connections between nodes. ANNs represent a paradigm for machine learning and can be used in
a variety of ways for image segmentation. The most widely applied use in medical imaging is as
a classifier, in which the weights are determined by using training data and the ANN is then used
to segment new data. ANNs can also be used in an unsupervised fashion as a clustering method,
as well as for deformable models. Because of the many interconnections used in a neural
network, spatial information can be easily incorporated into its classification procedures.
Although ANNs are inherently parallel, their processing is usually simulated on a standard serial
computer, thus reducing this potential computationaladvantage.

DeformableModels:
Deformable models are physically motivated, model-based techniques for delineating region
boundaries by using closed parametric curves or surfaces that deform under the influence of
internal and external forces. To delineate an object boundary in an image, a closed curve or
surface must first be placed near the desired boundary and then allowed to undergo an iterative
relaxation process. Internal forces are computed from within the curve or surface to keep it
smooth throughout the deformation. External forces are usually derived from the image to drive
the curve or surface toward the desired feature of interest.

Atlas based segmentation:

Atlas based segmentation approaches are the most frequently used and powerful approaches in
the field of medical image segmentation. In this, information on anatomy, shape, size, and
features of different, organs, soft tissues is compiled in the form of atlas or look up table (LUT).
Atlas guided approaches are similar to co-relation approaches and the plus point of atlas based
approaches is - it performs segmentation and classification in one go. Atlas based segmentation
approaches are among the third-generation algorithms. There are indications that certain atlas
based methods can compete with manual segmentations although atlas selection, atlas
registration procedure, and the manual tracing protocol used in atlas formation are factors that
can affect performance.[1] However, they face limitations in segmenting complex structure with
variable shape, size, and properties and expert knowledge is required in building the database.

2.4 Current research in cancer cell detection in images:

Proposed systems:

2.4.1 Image Acquisition

The first stage starts with taking a collection of CT scan images from the Database (ACSC).
Images are stored in MATLAB and displayed as a gray scale image. The lung CT images having
low noise when compared to scan image and MRI image. So we can take the CT images for
detecting the lungs. The main advantage of the computer tomography image having better clarity,
low noise and distortion For the experimental purpose 10 male images are examined his CT
scans were stored in database of images in JPEG/PNG image standards.

Image pre-processing

All the images have been undergoing several preprocessing process such as noise removal and
enhancement.
Noise Removal: Image denoising algorithms may be the mostly used in image processing. The
input image is a normal RGB image. The RGB image is converted into grey scale image because
the RGB format is not supported in Matlab. Then the grey scale image contains noises such as
white noise, salt and pepper noise etc White noise is one of the most common problems in image
processing. This can be removed by using filter from the extracted image.
Image Enhancement: Image enhancement defined as a way to improve the quality of image, so
that the resultant image is better than the original one, the process of improving the quality of a
digitally stored image by manipulating the image with MATLAB software. It is quite easy, for
example, to make an image lighter or darker, or to increase or decrease contrast. The aim of
image enhancement is to improve the visual appearance of an image, or to provide a better
transform representation for future automated image processing. Many images like medical
images, satellite images, aerial images and even real life photographs suffer from poor contrast
and noise. It is necessary to enhance the contrast and remove the noise to increase image quality.
The enhancement technique differs from one field to another according to its objective. In the
image enhancement stage we use Gabor filter enhancement technique.

2.4.2 Processing
This stage involves mainly segmentation which is explained as below:
Image Segmentation: In computer vision, segmentation refers to the process of partitioning a
digital image into multiple segments (sets of pixels, also known as superpixels).Image
segmentation is typically used to locate objects and boundaries (lines, curves, etc.) in images.
More precisely, image segmentation is the process of assigning a label to every pixel in an image
such that pixels with the same label share certain visual characteristics. The result of image
segmentation is a set of segments that collectively cover the entire image, or a set of contours
extracted from the image. Each of the pixels in a region is similar with respect to some
characteristic or computed property, such as color, intensity, texture. All image processing
operations generally aim at a better recognition of objects of interest, i.e., at finding suitable local
features that can be distinguished from other objects and from the background. The next step is
to check each individual pixel to see whether it belongs to an object of interest or not. This
operation is called segmentation and produces a binary image. A pixel has the value one if it
belongs to the object otherwise it is zero. After segmentation, it is known that which pixel
belongs to which object.

2.4.3 Post-Processing:
Post processing segmentation is done using following methods.
Thresholding approach: Thresholding is useful in discriminating foreground from the
background. By selecting an adequate threshold value T, the gray level image can be converted
to binary image. The binary image should contain all of the essential information about the
position and shape of the objects of interest (foreground). The advantage of obtaining first a
binary image is that it reduces the complexity of the data and simplifies the process of
recognition and classification. The most common way to convert a gray-level image to a binary
image is to select a single threshold value (T). Then all the gray level values below this T will be
classified as black (0), and those above T will be white (1). Otsus method using (gray thresh)
function Computes global image threshold. Otsus method is based on threshold selection by
statistical criteria. Otsu suggested minimizing the weighted sum of within-class variances of the
object and background pixels to establish an optimum threshold. Recall that minimization of
within-class variances is equivalent to maximization of between-class variance. This method
gives satisfactory results for bimodal histogram images.

2.4.4 Marker-Controlled Watershed Segmentation

In Marker-based watershed segmentation markers are used. A marker is a connected component
belonging to an image. The markers include the internal markers, associated with objects of
interest, and the external markers, associated with the background. Separating touching objects
in an image is one of the more difficult image processing operations. The water shed transform is
often applied to this problem. The marker based watershed segmentation can segment unique
boundaries from an image. The strength of watershed segmentation is that it produces a unique
solution for a particular image. The over-segmentation problem is also removed by marker
watershed segmentation. Generally, the watershed transform is computed on the gradient of the
original image. It possesses the number of advantages: it is a simple, intuitive method, it is fast
and can be parallelized and it produces a complete division of the image in separated regions
even if the contrast is poor.
2.4.5 Features Extraction
The Image features Extraction stage is very important in our working in image processing
techniques which using algorithms and techniques to detect and isolate various desired portions
or shapes (features) of an image. In the literature we found among the features used in the
diagnostic indicators. Two approaches to predict the probability of lung cancer presence first
approach is Binarization and the second is masking.
Binarization Approach: Binarization approach depends on the fact that the number of black
pixels is much greater than white pixels in normal lung images, so that the counting starts the
black pixels for normal and abnormal images to get an average that can be used later as a
threshold, if the number of the black pixels of a new image is greater that the threshold, then it
indicates that the image is normal, otherwise, if the number of the black pixels is less than the
threshold, it indicates that the image in abnormal.
Masking Approach: Masking approach depends on the fact that the masses are appeared as
white linked areas inside ROI (lungs), as they increase the percent of cancer presence increase.
The appearance of solid blue color indicates normal case while appearance of RGB masses
indicates the presence of cancer.
CHAPTER 3
NEED AND SIGNIFICANCE OF PROJECT
Image segmentation plays a crucial role in many medical-imaging applications, by automating or
facilitating the delineation of anatomical structures and other regions of interest. We present a
critical appraisal of the current status of semiautomated and automated methods for the
segmentation of anatomical medical images. Current segmentation approaches are then reviewed
with an emphasis on the advantages and disadvantages of these methods for medical imaging
applications.
Medical image plays an important role in the assist doctors in the diagnosis and treatment of
diseases. For the medical image, the further analysis and diagnosis of the target area is based on
image segmentation. In this project, image segmentation algorithms are divided into classical
image segmentation algorithms and segmentation methods combined with certain mathematical
tools, including threshold segmentation methods, image segmentation algorithms based on the
edge, image segmentation algorithms based on the region. Further the results are discussed.

3.1 Existing Systems

Segmentation Techniques:
Edge Detection Technique: The intensity data of an image only provides partial and uncertainly
information about the location of edges. Edge detection technique is finding pixel on the region
boundary.This method attempts to resolve image segmentation by detecting the edges or pixels
between different regions that have rapid transition in intensity are extracted and linked to form
closed object boundaries. The result is a binary image. It is also the filltering of the image with
operators such as Sobel, Prewitt, and Roberts gradient. This method is sensitive to noise and
must include a noise smoothing operation (Laplacian) before detecting the edges since isolated
noise pixels will also be detected as edges thereby compromising on efficient tracking of the
object boundary. This is accounted for by checking for the variance at zero-crossings.
Method Description: Based on the detection of discontinuity, normally tries to locate points
with more or less abrupt changes in gray level.
Advantages: Edge detection technique is the way in which human perceives objects and works
well for images having good contrast between regions.
Disadvantages:
1)Does not work well with images in which the edges are ill-defined or there are too many edges.
2)It is not a trivial job to produce a closed curve or boundary.
3)Less immune to noise than other techniques.

Thresholding Technique:Thresholding is the simplest Image Segmentation method. This

method is based on threshold value to convert the gray level image into a binary image. The key
of this method is to select the threshold value (or values when multiple-levels are selected).
Several popular methods are used in industry including the maximum entropy method, Otsu's
method (maximum variance), and et al k-means clustering can also be used. One of the simplest
approach of segmentation is based on the pixel values. The technique is to utilize the
thresholding based segementation which could help to simple region growing steps.
Thresholding algorithms can be selected manually as per a priori knowledge or automatically by
image information.
Method description: Requires that the histogram of an image has a number of peaks, each
correspond to a region.
Advantages: It does not need a prior information of the image.And it has less computational
complexity.
Disadvantages:
1)Does not work well for an image without any obvious peaks or with broad and flat valleys;
2)Does not consider the spatial details, so cannot guarantee that the segmented regions are
contiguous.

3.2 TOOLS REQUIRED

Matlab:
The name MATLAB stands for Matrix Laboratory. It was developed by Cleve Moler in the late
1970s. MATLAB was written originally to provide easy access to matrix software developed by
the LINPACK (linear system package) and EISPACK (Eigen system package) projects. It was
rewritten in C in 1983; The Math works was formed in 1984.
 MATLAB is a high-performance language for technical computing. It integrates computation,
visualization, and programming environment. Furthermore, MATLAB is a modern programming
language environment: it has sophisticated data structures, contains built-in editing and
debugging tools, and supports object-oriented programming. These factors make MATLAB an
excellent tool for teaching and research. MATLAB has many advantages compared to
conventional computer languages (e.g., C, FORTRAN) for solving technical problems.
MATLAB is an interactive system whose basic data element is an array that does not require
dimensioning. It is always recognized as one of the leading programs for linear algebra
It has powerful built-in routines that enable a very wide variety of computations. It also has easy
to use graphics commands that make the visualization of results immediately available. Specific
applications are collected in packages referred to as toolbox. There are toolboxes field
processing, symbolic computation, control theory, simulation, optimization, and several other
fields of applied science and engineering.
By default, matlab will open a console with a file browser and command history window along
with a window to input commands.
3.2.1 The major tools within or accessible from the MATLAB desktop are:
The Command Window: Command Window is used to enter data, run MATLAB code, and
display results. The Command Window prompt indicates that MATLAB is ready to accept input.
Enter statements at the prompt, which is also known as the command line.
The Command History: The Command History window displays a log of statements you ran in
the current and previous MATLAB sessions. The time and date for each session appear at the top
of the statements listed for that session. All entries remain until you delete them, or until the
command history file exceeds its maximum size of 200,000 bytes. When the file exceeds its
maximum size, MATLAB automatically deletes the oldest entries. Double-clicking re-executes a
command. Right-clicking brings up a menu of possible actions (e.g. delete from history)
The Workspace: The MATLAB workspace consists of the set of variables built up and stored in
memory during a MATLAB session. You add variables to the workspace by using functions,
running MATLAB code, and loading saved workspaces. Double-clicking on a name in the
Workspace Browser Window brings up the Array Editor, a spreadsheet-like window which
displays the current contents of the named variable or array and enables them to be changed
The Current Directory: It is used to see the list of folders.
 The Help Browser: Each MATLAB function has supporting documentation that includes
examples and describes the function inputs, outputs, and calling syntax.
The Start button: Brings up a menu of MATLAB tools, help files, settings etc (Windows
Start button)

Figure 3.1: The Graphical Interface To The MATLAB Workshop

1 MATLAB window.
2 Current Directory.
3 Command History.
4 Command Editor.
5 Workspace.
3.3 Strengths of Matlab:
MATLAB may behave as a calculator or as a programming language
MATLAB combine nicely calculation and graphic plotting.
MATLAB is relatively easy to learn.
MATLAB is interpreted (not compiled), errors are easy to fix.
MATLAB is optimized to be relatively fast when performing matrix operations
MATLAB does have some object-oriented elements
3.4 Weakness of Matlab:
MATLAB is not a general purpose programming language such as C, C++, or FORTRAN.
MATLAB is designed for scientific computing, and is not well suitable for other applications.
MATLAB is an interpreted language, slower than a compiled language such as C++.
MATLAB commands are specific for MATLAB usage. Most of them do not have a direct
equivalent with other programming language commands.
3.5 SUMMARY OF COMMANDS AND OPERATORS USED IN MATLAB
Table 3.1: Arithmetic Operators and Special Characters

Character Description
+ Addition

- Subtraction

* Multiplication (scalar and array)

/ Division (right)

^ Power or exponentiation

: Colon; creates vectors with equally spaced elements

; Semi-colon; suppresses display; ends row in array

, Comma; separates array subscripts

% Percent; denotes a comment; specifies output format

Single quote; creates string; specifies matrix transpose

= Assignment operator

() Parentheses; encloses elements of arrays and input arguments

[] Brackets; encloses matrix elements and output arguments

 Table 3.2: Array Operators

Character Description

.* Array multiplication

./ Array (right) division

.^ Array power

.\ Array (left) division

. Array (non-conjugated) transpose

Table 3.3: Relational And Logical Operators

Character Description

< Less than

<= Less than or equal to

> Greater than

>= Greater than or equal to

== Equal to

~= Not equal to
& Logical or element-wise AND

| Logical or element-wise OR

&& Short-circuit AND

|| Short-circuit OR

Table 3.4: List of Matlab Commands Used

Command Description

Clc Clear the Command Window

clear all Removes all variables from the workspace.

close all Close all the windows.

help Display help for MATLAB functions.

size Size of matrix.

length Length of vector

imread IMREAD Read image from graphics file.

A = imread (FILENAME.FMT); reads a grayscale or color image
from the file specified by the string FILENAME. If the file is not in
the current directory, or in a directory on the MATLAB path, specify
the full pathname.
The text string FMT specifies the format of the file by its standard file
extension.
im2double IM2DOUBLE Convert image to double precision.
A = im2double (image); takes an image as input, and returns an image
of class double. If the input image is of class double, the output image
is dentical to it. If the input image is not double, IM2DOUBLE returns
the equivalent image of class double, rescaling or offsetting the data as
necessary.

figure FIGURE, by itself, creates a new figure window, and returns its
handle.
figure (H); makes H the current figure, forces it to become visible, and
raises it above all other figures on the screen. If Figure H does not
exist, then H is an integer, a new figure is created with handle H.

imshow IMSHOW Display image in Handle Graphics figure.

imshow (FILENAME); displays the image stored in the graphics file
FILENAME. The file must contain an image that can be read by
IMREAD. IMSHOW calls IMREAD or to read the image from the
file, but does not store the image data in the MATLAB workspace. If
the file contains multiple images, the first one will be displayed. The
file must be in the

title title ('text') adds text at the top of the current axis.

xlabel xlabel ('text') adds text beside the X-axis on the current axis.

ylabel ylabel ('text') adds text beside the Y-axis on the current axis.

legend legend (string1,string2,string3, ...) puts a legend on the current plot

using the specified strings as labels. LEGEND works on line graphs,
bar graphs, pie graphs, ribbon plots, etc. You can label any solid-
 colored patch or surface object.

plot plot(X,Y) plots vector Y versus vector X. If X or Y is a matrix, then

the vector is plotted versus the rows or columns of the matrix,
whichever line up. If X is a scalar and Y is a vector, disconnected line
objects are created and plotted as discrete points vertically at X.

rgb2gray Image = rgb2gray(rgb_img) converts RGB images to grayscale by

eliminating the hue and saturation information while retaining the
luminance.

graycomatrix GLCMS = graycomatrix (I) analyzes pairs of horizontally adjacent

pixels in a scaled version of I. If I a binary image, it is scaled to 2
levels. If I is an intensity image, it is scaled to 8 levels. In this case,
there are 8 x 8 = 64 possible ordered combinations of values for each
pixel pair. GRAYCOMATRIX accumulates the total occurrence of
each such combination, producing an 8-by-8 output array, GLCMS.
The row and column subscripts in GLCMS correspond respectively to
the first and second (scaled) pixel-pair values.

histeq histeq enhances the contrast of images by transforming the values in

an intensity image, or the values in the color map of an indexed
image, so that the histogram of the output image approximately
matches a specified histogram.

adapthisteq adapthisteq enhances the contrast of images by transforming the

values in the intensity image I. Unlike HISTEQ, it operates on small
data regions (tiles), rather than the entire image. Each tile's contrast is
enhanced, so that the histogram of the output region approximately
matches the specified histogram. The neighboring tiles are then
combined using bilinear interpolation in order to eliminate
 artificially induced boundaries. The contrast, especially in
homogeneous areas, can be limited in order to avoid amplifying the
noise which might be present in the image.

CHAPTER 4
OBJECTIVE
Medical image processing is an exciting and active field of research , where disciplines such as
Engineering, Computer Science, Physics, Biology and Medicine interdisciplinarily cooperate in
order to improve healthcare. Most frequently, medical images are the basis of diagnostics,
treatment planning, and treatment, but medical images are likewise important for medical
education, research and epidemiology.

Segmentation is the process dividing an image into regions with similar properties such as gray
level, color, texture, brightness, and contrast.[79] The role of segmentation is to subdivide the
objects in an image; in case of medical image segmentation the objective is to:

Study anatomical structure

Identify Region of Interest i.e. locate tumor, lesion and other abnormalities

Measure tissue volume to measure growth of tumor (also decrease in size of tumor with
treatment)

Help in treatment planning prior to radiation therapy; in radiation dose calculation

To automate the process so that large number of cases can be handled with the same
accuracy i.e. the results are not affected as a result of fatigue, data overload or missing
manual steps.

To achieve fast and accurate results. Very high-speed computers are, now, available at
modest costs, speeding up computer-based processing in the medical field.

To support faster communication, wherein patient care can be extended to remote areas
using information technology.
CHAPTER 5

IMPLEMENTATION

This section shows how to detect a cell using edge detection and basic morphology. An object
can be easily detected in an image if the object has sufficient contrast from the background. In
this project, the cells are prostate cancer cells.

Edge Detection:

Edge detection is the name for a set of mathematical methods which aim at identifying points in
a digital image at which the image brightness changes sharply or, more formally, has
discontinuities. The points at which image brightness changes sharply are typically organized
into a set of curved line segments termed edges. The same problem of finding discontinuities in
1D signals is known as step detection and the problem of finding signal discontinuities over time
is known as change detection. Edge detection is a fundamental tool in image processing, machine
vision and computer vision, particularly in the areas of feature detection and feature extraction.

Thresholding and Linking:

Once we have computed a measure of edge strength (typically the gradient magnitude), the next
stage is to apply a threshold, to decide whether edges are present or not at an image point. The
lower the threshold, the more edges will be detected, and the result will be increasingly
susceptible to noise and detecting edges of irrelevant features in the image. Conversely a high
threshold may miss subtle edges, or result in fragmented edges.If the edge thresholding is applied
to just the gradient magnitude image, the resulting edges will in general be thick and some type
of edge thinning post-processing is necessary. For edges detected with non-maximum
suppression however, the edge curves are thin by definition and the edge pixels can be linked
into edge polygon by an edge linking (edge tracking) procedure. On a discrete grid, the non-
maximum suppression stage can be implemented by estimating the gradient direction using first-
order derivatives, then rounding off the gradient direction to multiples of 45 degrees, and finally
comparing the values of the gradient magnitude in the estimated gradient direction.

Morphological Operations
Morphological processing is constructed with operations on sets of pixels. Binary morphology
uses only set membership and is indifferent to the value, such as gray level or color, of a pixel. It
relies on the ordering of pixels in an image and many times is applied to binary and gray scale
images. Through processes such as erosion, dilation, opening and closing, Binary images can be
modified to the user's specifications. Binary images are images whose pixels have only two
possible intensity values. They are normally displayed as black and white. Numerically, the two
values are often 0 for black, and either 1 or 255 for white. Binary images are often produced by
thresholding a gray scale or color image in order to separate an object in the image from the
background. The color of the object (usually white) is referred to as the foreground color. The
rest (usually black) is referred to as the background color.

Morphological Operators:
After converting the image in the binary format some morphological operations are applied on
the converted binary image. The purpose of the morphological operators is to separate the
tumour part of the image. The portion of the tumour in the image is visible as white color which
has the highest intensity then other regions of the image. Some of the commands used in the
morphing are strel which is used for creating morphological structuring element, imerode which
is used to erode or shrink an image and imdilate which is used to for dilating i.e. expanding an
image[7].After segmentation and thresholding some percent of noise will be there, in order to
remove this noise two important morphological operations have been used: opening and
closing.
These morphological operations basically used are
1.Dilation: Dilation is generally used for thickening of the object.
2.Erosion: Erosion is generally used for thinning of the object.
3.Opening: Opening is generally used for smoothening the contour of an object or elimination of
thin protrusions.
4.Closing: Closing is generally used for filling gaps between close objects.
Smoothing
Images taken from a camera will contain some amount of noise. As noise canmislead the result
in finding edges, we have to reduce the noise. Therefore theimage is first smoothed [16] by
applying a Gaussian filter.
We have implemented Gaussian filtering with specific kernel size (N) and Gaussianenvelope
parameter _. The Gaussian filter mask of NxN size is generated byinvoking following function.
private void GenerateGaussianKernel(int N, float S, out int Weight). The function
GenerateGaussianKernel takes kernel size ( parameterN) and theenvelope parameter _
(parameter S) as input to generate the NxN kernel values.
Following subroutine removes noise by Gaussian filtering private int[,] GaussianFilter(int[,]
Data). The function GaussianFilter multiplies each pixel in the image by the kernel
generated. It returns the smoothed image in a two dimensional array.

Finding Gradients
The principle of Canny algorithm is based on finding edges where the intensityof the grey image
changes to maximum value. These points are marked bydetermining the gradient of the image.
Gradient for each pixel is calculated byapplying Sobel operator. For each pixel, partial gradient
towards x and y directionis determined respectively by applying the kernels.By applying
Pythagoras law, we have calculated themagnitude of gradient (strength of the edge) for each
point. The edges have been marked where the gradients of the image have large magnitudes.
SobelGx and Gy masks are used to generate partial derivatives @x and@y (partial spatial
derivatives) of the image. Spatial gradient value (d f (x; y)) isobtained from this partial
derivatives. The following function implements differentiationusing Sobel filter mask.
private float[,] Differentiate(int[,] Data, int[,] Filter)
The function takes the Gray image as input. Color value of Gray image isstored in a two
dimensional array. The image array is smoothed first. Later, aderivative of the image is
performed by calling the function Differentiate. Thisfunction returns gradient image as a two
dimensional array.
SOURCE CODE
Step 1: Read Image
Read in the cell.tif image, which is an image of a prostate cancer cell.
I = imread('cell.tif');
figure, imshow(I), title('original image');
text(size(I,2),size(I,1)+15, ...
'Image courtesy of Alan Partin', ...
'FontSize',7,'HorizontalAlignment','right');
text(size(I,2),size(I,1)+25, ....
'Johns Hopkins University', ...
'FontSize',7,'HorizontalAlignment','right');

Step 2: Detect Entire Cell

Two cells are present in this image, but only one cell can be seen in its entirety. We will detect
this cell. Another word for object detection is segmentation. The object to be segmented differs
greatly in contrast from the background image. Changes in contrast can be detected by operators
that calculate the gradient of an image. The gradient image can be calculated and a threshold can
be applied to create a binary mask containing the segmented cell. First, we use edge and the
Sobel operator to calculate the threshold value. We then tune the threshold value and
useedge again to obtain a binary mask that contains the segmented cell.
[~, threshold] = edge(I, 'sobel');
fudgeFactor = .5;
BWs = edge(I,'sobel', threshold * fudgeFactor);
figure, imshow(BWs), title('binary gradient mask');

Step 3: Dilate the Image

The binary gradient mask shows lines of high contrast in the image. These lines do not quite
delineate the outline of the object of interest. Compared to the original image, you can see gaps
in the lines surrounding the object in the gradient mask. These linear gaps will disappear if the
Sobel image is dilated using linear structuring elements, which we can create with
the strel function.
se90 = strel('line', 3, 90);
se0 = strel('line', 3, 0);
The binary gradient mask is dilated using the vertical structuring element followed by the
horizontal structuring element. The imdilate function dilates the image.
BWsdil = imdilate(BWs, [se90 se0]);
figure, imshow(BWsdil), title('dilated gradient mask');
Step 4: Fill Interior Gaps
The dilated gradient mask shows the outline of the cell quite nicely, but there are still holes in the
interior of the cell. To fill these holes we use the imfill function.
BWdfill = imfill(BWsdil, 'holes');
figure, imshow(BWdfill);
title('binary image with filled holes');

Step 5: Remove Connected Objects on Border

The cell of interest has been successfully segmented, but it is not the only object that has been
found. Any objects that are connected to the border of the image can be removed using the
imclearborder function. The connectivity in the imclearborder function was set to 4 to remove
diagonal connections.
BWnobord = imclearborder(BWdfill, 4);
figure, imshow(BWnobord), title('cleared border image');

Step 6: Smoothen the Object

Finally, in order to make the segmented object look natural, we smoothen the object by eroding
the image twice with a diamond structuring element. We create the diamond structuring element
using the strel function.
seD = strel('diamond',1);
BWfinal = imerode(BWnobord,seD);
BWfinal = imerode(BWfinal,seD);
figure, imshow(BWfinal), title('segmented image');

An alternate method for displaying the segmented object would be to place an outline around the
segmented cell. The outline is created by the bwperim function.
BWoutline = bwperim(BWfinal);
Segout = I;
Segout(BWoutline) = 255;
figure, imshow(Segout), title('outlined original image');

Conclusions
Segmentation techniques based on gray level techniques such as thresholding, and region based
techniques are the simplest techniques and find limited applications. However, their performance
can be improved by integrating them with artificial intelligence techniques. Techniques based on
textural features utilizing atlas or look-up-table have excellent results on medical image
segmentation. However, they need expert knowledge in building the atlas. The limitation of atlas
based technique is that under certain circumstances it becomes difficult to correctly select and
label data; has difficulties in segmenting complex structure with variable shape, size, and
properties. In such situations it is better to use unsupervised methods such as fuzzy-c-means
algorithm.
References:
1. Research project on image contrast enhancement methods dept. of radio
communication and video technology; university of SOFIA.
2. Efficient algorithm for contrast enhancement of natural images ShyamLal and Mahesh
Chandra; department of ECE, Moradabad Institute of Technology, India.
3. Review of various image contrast enhancement techniques Vijay A. Kotkar, Sanjay S.
Ghardi; research scholar, dept of CSE, Maharastra, India.
4. Contrast limited adaptive histogram equalization for qualitative enhancement Neethu
M. Sasi, V.K Jayasree; Govt. Model Engg. College, Cochin University of Science &
Technology, Thirkkatkara, Kerela, India.
5. Region based contrast limited adaptive HE Sonia Goyal, Seema ,YCOE; Punjabi
University Campus; Talwandi Sabo, Punjab, India.
6. Different histogram equalization based contrast enhancement techniques Shefali Gupta,
YadwinderKaur; Chandigarh group of colleges, Gharuan, Mohali, India.
7. A comparative analysis of histogram equalization based techniques for contrast
enhancement and brightness preserving Raju A. Dwarakish, G.S and Venkant Reddy;
National Institute of Technology, Karnataka, India.
8. Adaptive image contrast enhancement using generalizations of histogram equalization
J. Alex Stark, Cambridge University U.K.
9. Comparison of histogram equalization techniques for image enhancement of grayscale
images Dinesh Sonkar, M.R. Parsai, dept. of ECE, Jabalpur Engg. College, Jabalpur,
M.P. India.