White Blood Cells Identification and Counting From Microscopic Blood Image
White Blood Cells Identification and Counting From Microscopic Blood Image
White Blood Cells Identification and Counting From Microscopic Blood Image
CHAPTER1
INTRODUCTION
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INTRODUCTION
1.1 IMAGE
An image is a two-dimensional picture, which has a similar appearance to some
subject usually a physical object or a person.
Image is a two-dimensional, such as a photograph, screen display, and as well as a
three-dimensional, such as a statue. They may be captured by optical devicessuch as
cameras, mirrors, lenses, telescopes, microscopes, etc. and natural objects and phenomena,
such as the human eye or water surfaces.
The word image is also used in the broader sense of any two-dimensional figure such
as a map, a graph, a pie chart, or an abstract painting. In this wider sense, images can also be
rendered manually, such as by drawing, painting, carving, rendered automatically by printing
or computer graphics technology, or developed by a combination of methods, especially in a
pseudo-photograph.
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example, an image recorded by a 12 MP camera; since each pixel uses 3 bytes to record true
color, the uncompressed image would occupy 36,000,000 bytes of memory, a great amount
of digital storage for one image, given that cameras must record and store many images to be
practical. Faced with large file sizes, both within the camera and a storage disc, image file
formats were developed to store such large images.
1.1.3RASTER FORMATS
These formats store images as bitmaps (also known as pixmaps).
JPEG/JFIF
JPEG (Joint Photographic Experts Group) is a compression method. JPEG
compressed images are usually stored in the JFIF (JPEG File Interchange Format) file format.
JPEG compression is lossy compression. Nearly every digital camera can save images in the
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JPEG/JFIF format, which supports 8 bits per color (red, green, blue) for a 24-bit total,
producing relatively small files. Photographic images may be better stored in a lossless nonJPEG format if they will be re-edited, or if small "artifacts" are unacceptable. The JPEG/JFIF
format also is used as the image compression algorithm in many Adobe PDF files.
EXIF
The EXIF (Exchangeable image file format) format is a file standard similar to the JFIF
format with TIFF extensions. It is incorporated in the JPEG writing software used in most
cameras. Its purpose is to record and to standardize the exchange of images with image
metadata between digital cameras and editing and viewing software. The metadata are
recorded for individual images and include such things as camera settings, time and date,
shutter speed, exposure, image size, compression, name of camera, color information, etc.
When images are viewed or edited by image editing software, all of this image information
can be display.
TIFF
The TIFF (Tagged Image File Format) format is a flexible format that normally saves
8 bits or 16 bits per color (red, green, blue) for 24-bit and 48-bit totals, respectively, usually
using either the TIFF or TIF filename extension. TIFFs are lossy and lossless. Some offer
relatively good lossless compression for bi-level (black & white) images. Some digital
cameras can save in TIFF format, using the LZW compression algorithm for lossless storage.
TIFF image format is not widely supported by web browsers. TIFF remains widely accepted
as a photograph file standard in the printing business. TIFF can handle device-specific color
spaces, such as the CMYK defined by a particular set of printing press inks.
PNG
The PNG (Portable Network Graphics) file format was created as the free, opensource successor to the GIF. The PNG file format supports true color (16 million colors)
while the GIF supports only 256 colors. The PNG file excels when the image has large,
uniformly colored areas. The lossless PNG format is best suited for editing pictures, and the
lossy formats, like JPG, are best for the final distribution of photographic images, because
JPG files are smaller than PNG files. PNG, an extensible file format for the lossless, portable,
well-compressed storage of raster images. PNG provides a patent-free replacement for GIF
and can also replace many common uses of TIFF. Indexed-color, grayscale, and true color
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images are supported, plus an optional alpha channel. PNG is designed to work well in online
viewing applications, such as the World Wide Web. PNG is robust, providing both full file
integrity checking and simple detection of common transmission errors.
GIF
GIF (Graphics Interchange Format) is limited to an 8-bit palette, or 256 colors. This
makes the GIF format suitable for storing graphics with relatively few colors such as simple
diagrams, shapes, logos and cartoon style images. The GIF format supports animation and is
still widely used to provide image animation effects. It also uses a lossless compression that
is more effective when large areas have a single color, and ineffective for detailed images or
dithered images.
BMP
The BMP file format (Windows bitmap) handles graphics files within the Microsoft
Windows OS. Typically, BMP files are uncompressed, hence they are large. The advantage is
their simplicity and wide acceptance in Windows programs.
CGM
CGM (Computer Graphics Metafile) is a file format for 2D vector graphics, raster
graphics, and text. All graphical elements can be specified in a textual source file
that can be compiled into a binary file or one of two text representations. CGM
provides a means of graphics data interchange for computer representation of 2D
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graphical information independent from any particular application, system,
platform, or device.
SVG
SVG (Scalable Vector Graphics) is an open standard created and developed by the
World Wide Web Consortium to address the need for a versatile, scriptable and all purpose
vector format for the web and otherwise. The SVG format does not have a compression
scheme of its own, but due to the textual nature of XML, an SVG graphic can be compressed
using a program such as gzip.
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Enhancement, on the other hand, is based on human subjective preferences regarding
what constitutes a good enhancement result. For example, contrast stretching is considered
an enhancement technique because it is based primarily on the pleasing aspects it might
present to the viewer, where as removal of image blur by applying a deblurring function is
considered a restoration technique.
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1.2.6 COMPRESSION
Compression, as the name implies, deals with techniques for reducing the storage
required saving an image, or the bandwidth required for transmitting it. Although storage
technology has improved significantly over the past decade, the same cannot be said for
transmission capacity. This is true particularly in uses of the Internet, which are characterized
by significant pictorial content. Image compression is familiar to most users of computers in
the form of image file extensions, such as the jpg file extension used in the JPEG (Joint
Photographic Experts Group) image compression standard.
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1.2.8 SEGMENTATION
Segmentation procedures partition an image into its constituent parts or objects. In general,
autonomous segmentation is one of the most difficult tasks in digital image processing. A
rugged segmentation procedure brings the process a long way toward successful solution of
imaging problems that require objects to be identified individually.
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Network
Image displays
Hard copy
computer
Mass storage
Specialized image
processing
hardware
Image processing
software
Image sensor
Problem domain
Figure 1.13: Components Of An Image Processing System
Although large-scale image processing systems still are being sold for massive
imaging applications, such as processing of satellite images, the trend continues toward
miniaturizing and blending of general-purpose small computers with specialized image
processing hardware. Figure 1.13 shows the basic components comprising a typical generalpurpose system used for digital image processing. The function of each component is
discussed in the following paragraphs, starting with image sensing.
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IMAGE SENSORS
With reference to sensing, two elements are required to acquire digital images. The
first is a physical device that is sensitive to the energy radiated by the object we wish to
image. The second, called a digitizer, is a device for converting the output of the physical
sensing device into digital form. For instance, in a digital video camera, the sensors produce
an electrical output proportional to light intensity. The digitizer converts these outputs to
digital data.
COMPUTER
The computer in an image processing system is a general-purpose computer and can
range from a PC to a supercomputer. In dedicated applications, sometimes specially designed
computers are used to achieve a required level of performance, but our interest here is on
general-purpose image processing systems. In these systems, almost any well-equipped PCtype machine is suitable for offline image processing tasks.
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MASS STORAGE
Mass storage capability is a must in image processing applications. An image of size
1024*1024 pixels, in which the intensity of each pixel is an 8-bit quantity, requires one
megabyte of storage space if the image is not compressed. When dealing with thousands, or
even millions, of images, providing adequate storage in an image processing system can be a
challenge.
Digital storage for image processing applications fall into three principal categories:
(1) short-term storage for use during processing, (2) on-line storage for relatively fast recall,
and (3) archival storage, characterized by infrequent access. Storage is measured in bytes
(eight bits), Kbytes (one thousand bytes), Mbytes (one million bytes), Gbytes (meaning giga,
or one billion, bytes), and Tbytes (meaning tera, or one trillion, bytes).
One method of providing short-term storage is computer memory. Another is by
specialized boards, called frame buffers that store one or more images and can be accessed
rapidly, usually at video rates. The latter method allows virtually instantaneous image zoom,
as well as scroll (vertical shifts) and pan (horizontal shifts). Frame buffers usually are housed
in the specialized image processing hardware unit.
Online storage generally takes the form of magnetic disks or optical-media storage.
The key factor characterizing on-line storage is frequent access to the stored data. Finally,
archival storage is characterized by massive storage requirements but infrequent need for
access. Magnetic tapes and optical disks housed in jukeboxes are the usual media for
archival applications.
IMAGE DISPLAYS
Image displays in use today are mainly color (preferably flat screen) TV monitors.
Monitors are driven by the outputs of image and graphics display cards that are an integral
part of the computer system. Seldom are there requirements for image display applications
that cannot be met by display cards available commercially as part of the computer system. In
some cases, it is necessary to have stereo displays, and these are implemented in the form of
headgear containing two small displays embedded in goggles worn by the user.
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HARD COPY
Hardcopy devices for recording images include laser printers, film cameras, heatsensitive devices, inkjet units, and digital units, such as optical and CD-ROM disks. Film
provides the highest possible resolution, but paper is the obvious medium of choice for
written material. For presentations, images are displayed on film transparencies or in a digital
medium if image projection equipment is used. The latter approach is gaining acceptance as
the standard for image presentations.
NETWORK
Networking is almost a default function in any computer system in use today. Because
of the large amount of data inherent in image processing applications, the key consideration
in image transmission is bandwidth. In dedicated networks, this typically is not a problem,
but communications with remote sites via the Internet are not always as efficient. Fortunately,
this situation is improving quickly as a result of optical fiber and other broadband
technologies.
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The use of image processing techniques has grown rapidly in the recent years. These
techniques help to count the cells in the human blood and, at the same time, provide
information on the cells morphology. Unfortunately, there are not standard techniques for the
analysis and processing of the images valid for each type of them, but the processing must be
adapted to the context. Regarding the microscopic images, processing techniques vary
depending on the type of blood cells to be analyzed. Our work focuses on the analysis of
white blood cells or WBCs. A typical blood image usually shows four components: plasma,
red blood cellsor erythrocytes, white blood cells or leukocytes, and platelets. The most
represented cells in the blood are red blood cells and white blood cells. Leukocytes are easily
identifiable, as their nucleus appears darker than the background.
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CHAPTER2
LITERATURE SURVEY
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LITERATURE SURVEY
2.1 A NEW WHITE BLOOD CELL SEGMENTATION USING MEAN SHIFT
FILTER AND REGION GROWING ALGORITHM
The differential counting of white blood cells (WBCs) provides invaluable
information to hematologist for diagnosis and treatment of many diseases. However,
manually counting the WBCs is a time consuming and susceptible to error procedure. Then
an automatic and efficient system seems necessary and helpful. In the automatic process, the
segmentation of WBC image is one of the most important stages. In this paper, we propose a
new algorithm to segment the image.
The algorithm consists of two tasks. Firstly, the Mean Shift (MS) filter is used to remove
noise. Secondly, we propose a simple but effective region growing algorithm to segment the
image. The proposed algorithm was tested in RGB color space and CIE L*a*b* color space,
and the experimental results show the excellent performance in both color spaces.
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samples, specifically designed for the evaluation and the comparison of algorithms for
segmentation and classification. For each image in the dataset, the classification of the cells is
given, as well as a specific set of figures of merits to fairly compare the performances of
different algorithms. This initiative aims to offer a new test tool to the image processing and
pattern matching communities, direct to stimulating new studies in this important field of
research.
2.4. ROBUST RECOGNITION OF WHITE BLOOD CELL IMAGES
The objective of this work is to investigate the white blood cell (WBC) image
recognition problem at all stages. A robust and effective method for automatic WBC
differentiation, based on both statistical pattern recognition and neural net approaches, is
presented.
We demonstrate well-evaluated results ranging from image scene segmentation techniques to
recognition details. Recognition accuracy on the test set of 662 images of five WBC types
obtained by different imaging systems from 22 bloodstains is not less than 98%.
2.6. DRAWBACKS
Slowness of the analysis and
A nonstandard accuracy,
Dependent on the operator skills.
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CHAPTER 3
PROPOSED METHOD
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PROPOSED METHOD
3.1 OBJECTIVE OF PRESENT STUDY
This technique presents a complete and fully automatic method for white blood cells
identification from microscopic images. The proposed method firstly individuates white
blood cells from which, subsequently, nucleus and cytoplasm are extracted.
3.2 INTRODUCTION
The proposed method, differently from other methods in the literature, does not
present separate steps of pre-processing and segmentation, but uses methods of preprocessing inserted between the various stages of the segmentation, in order to make the
latter more simple and more robust.
The identification of the leukocytes is carried out in the first stage, this requires also
identifying and removing the image background, in order to make the identification of
leukocytes more efficient. The result consists of a binary image showing only leukocytes.
The second step is the identification of leukocytes groups and produces a binary image
showing the individual leukocytes and a binary image showing the adjacent leukocytes.
The third phase takes care of separating the adjacent leukocytes and removing the elements
localized on the border of the image.
The fourth phase proceeds with a shape control, through which all the abnormal components
are removed from the image.
The fifth and last phase deal with the selection of the nucleus and the cytoplasm of each
leukocyte.
The whole process can be schematized as showed in Figure 3.1.
Proposed method entirely will be done in above five steps as shown above , the entire process
can be done using the basic image processing techniques like converting to gray images,
enhancement techniques, thresholding ,morphological operations like region growing using
opening and dilation techniques.
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In the proposed scheme ,the simple five steps are enough. In the first step, leukocytes
identification
identified . In the third step , the adjacent leukocytes are get separated. Fourth stage includes
the shape control and abnormal components are removed. Finally, the selection of nucleus
and cytoplasm of each leukocytes is done.
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3.3 METHODOLOGY
A. Background Identification
Since that the images captured at the microscope suffer from uneven lighting, it becomes
necessary to remove the background because the segmentation methods based on threshold
may suffer heavily for this problem. Some methods for background extraction are present in
literature, but they use a collection of images captured with the same camera and the same
microscope for estimation of the pixels belonging to the background, while others have a
very high computational cost, not necessary for this application. The proposed approach
involves the use of an automatic threshold to the original image in gray level (or along the
green component, in this case the image chosen is the one presenting the greatest difference
between minimum and maximum intensity level). There are many threshold techniques
available in literature. Here, we use the threshold value based on triangle method or Zack
algorithm. The triangle method is applied to the image histogram, constructing a straight line
that connects the highest histogram value h[ ] and the lowest histogram value h[ ],
where and indicatethe values of the gray levels where the histogram h[x] reaches
its maximum and minimum, respectively. The distance between the marked line and the
histogram values between and is then calculated.
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leukocytes identification also the red blood cells will be removed. Background removal can
be performed later (as described in the proposed method) or directly in this phase, starting
from the original image of the blood sample in the RGB color space or from the gray level
image. In both cases the background removal can be performed by simple arithmetical
operations.
B. Leukocytes Identification
In many methods present in the literature the idea is to identify firstly the nucleus
which are more prominent than other components and then the entire membrane, for example
by region growing. In the proposed method instead, the membrane is detected firstly, inorder
to deal the subsequent separation of the adjacent cells more accurately. The white blood cells
identification wasmade possible thanks to the conversion into the CMYK color model. In
fact, we have observed that leukocytes are more contrasted in the Y component of CMYK
color model, this is because the yellow color is present in all the elements of the image except
in leukocytes, where it is practically absent.
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C. Background Removal
The operation performed to identify more accurately the white blood cells is the same
previously seen for the background removal .
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Figure.3.8: Two Original Blood Sample Sub-images And Their Respective Watershed
Results
For this reason it is necessary a second step to refine the contours extracted through
watershed transform. Then, althea pixels of the component under examination which are
located at a distance not greater than a predetermined value from the watershed line
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concerned, are taken in consideration. These pixels are then used to derive the deepest
concavity for which the line of exact separation will have to pass. Therefore, by exploiting
the information of the points of concavity and the information related to the points of
maximum image in gray tones, it is possible to obtain a cutting line that best fits the contour
of the leukocytes, as we can see in Figure. 11.
F. Image Cleaning
The image cleaning requires the removal of all the leukocytes located along the
border of the image and of all abnormal components (that are not leukocytes), in order to
avoid errors in the later stages of the analysis process. The cleaning of the image border is a
simple operation, while the removal of abnormal components is a more complex process.
To do this it is necessary to first determine the number of leukocytes present in the
image. For each of them it is then calculated the size of the area and the size of the convex
area. The size of the area is used to calculate the mean area, necessary to determine and
eliminate the components with irregular dimensions. For example, a very small area might
indicate the presence of artifacts not removed adequately, on the other side, a very large area
may indicate the presence of adjacent leukocytes not separated adequately.
Area and convex area are then used in combination for the calculation of the solidity
value. All objects with a solidity value less thana predetermined threshold are discarded. In
fact, even in thiscase, a value of solidity less than the threshold value indicates the presence
of artifacts not removed adequately.
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Figure.3.11: Gray level And Binary Sub-image Of Individual Leukocytes. Border Cleaning
From Binary Image. Gray level Sub-image With
Superimposed Border. Gray level Sub-image With External Leukocyte Image Cropped
This step can be simplified performing an image crop using the bounding box size
that is the smallest rectangle that completely contains the connected component, with the aim
to have a single leukocyte for sub-image. A border cleaning operation is again necessary in
order to preserve only the white blood cell in question.
Since by definition, leukocytes nucleus is internal to the membrane, it is possible to
perform further simplification, through the crop of the entire portion of the image outside the
leukocyte in question . This procedure allows a more robust nucleus selection, because it
excludes completely artifacts from the selection. Nucleus selection approach takes advantage
from Csekes observations, who found that white blood cells nuclei are more in contrast in
the green component of the RGB color space.
Threshold operation using Otsu in this color space, however, does not produce clean
results, especially with the presence of granulocytes, whose granules are selected erroneously
as part of the nucleus.
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Figure. 3.12: Images Of The G Component In The RGB space And a* Component In The
CIE Lab Space. Binary Image After Nucleus Selection. Binary Image After Cytoplasm
Selection
For this the binary image obtained from the green component, is combined with the
binary image, obtained from the a* component of the CIEL a, b color space, again through a
threshold operation.
The mask obtained makes it possible to extract clearly the leukocytes nucleus. At the
end, to obtain the cytoplasm you just have to perform a subtraction operation between the
binary image containing the whole leukocyte and the image containing only the nucleus.
Finally, the nuclei and cytoplasm of leukocytes is identified .Then after performing the
identification of leukocytes by using thresholding, and region growing like morphological
operations, then the counting of leukocytes is done .
Here when compared to existing methodologies, proposed method identifies and counts the
no. of leukocytes is done efficiently.
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CHAPTER4
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4.1 RESULTS
In this picture , the white blood cells count is 3. In this the count is very less
in the taken blood sample
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In this third sample of blood sample of image, the white blood cell count is
123.
4.2 APPLICATIONS
1.WBC count is useful in detecting and treating leukemia.
2.WBC count places a major role in detecting bone marrow diseases.
3.WBC count is the basic measure to be calculated to detect any disease attacked on
human body.
4.Leukocytes count is useful in detecting zica virus.
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CHAPTER 5
CONCLUSION
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CONCLUSION
In this work it has been proposed an innovative method for a completely automatic
identification of leukocytes from microscopic images, in order to provide an automated
procedure as support for medical activity. The results obtained show that the proposed
method is able to identify in a robust way the white blood cells present in the image,
separating the agglomerates of cells and selecting the nucleus and the cytoplasm.
Further developments of the proposed method could affect the phase of separation of
adjacent leukocytes, which, despite producing robust results, require a better tool than the
current one, for both the identification of groups of leukocytes (even artifacts and strips of
dye present in the image are identified as adjacent white blood cells), and for the separation
itself, which generates incorrect results with the presence of holes to the whole leukocyte.
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45) J. M. S. Prewitt and M. L. Mendelsohn, \A general approach to image analysis by
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BIODATA
1.
2.
3.
4.
Name
Regd. No
Fathers Name
Address
:
:
:
:
Phone
Email
:
:
Name
Regd. No
Fathers Name
Address
:
:
:
:
Phone
Email
:
:
Name
Regd. No
Fathers Name
Address
:
:
:
:
M.MADHAVARAO
12JE1A0431
M.VENKATESWARLU
Kunduruvaripalem(V), Muppalla (M)
Guntur (DT), A.P.
Phone
9000143614
madhavmg143@gmail.com
Name
Regd. No
Fathers Name
Address
:
:
:
:
Phone
Email
:
:
V.NARENDRA BABU
12JE1A0450
V.NAGESWARA RAO
Karumanchi(V),Savalyapuram (M)
Guntur (DT), A.P.
9160022847
v50narendra@gmail.com
Department of ECE
J.MANO JYOTIRMAI
12JE1A0434
J.NAGESWARA RAO
Gurijepalli (v), Santamaguluru(M)
Prakasam (DT), A.P.
9666014683
manojyoti434@gmail.com
K.DIVYA
12JE1A0414
K.SRINIVASARAO
Boppudi(V), Chilakaluripet (M)
Guntur (DT), A.P.
9618894627
divyakatru414@gmail.com
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