International Journal of Emerging Trends & Technology in Computer Science (IJETTCS)

Web Site: www.ijettcs.org Email: editor@ijettcs.org, editorijettcs@gmail.com Volume 2, Issue 3, May June 2013 ISSN 2278-6856

White Blood Cells Segmentation and Classification to Detect Acute Leukemia

Ms. Minal D. Joshi1, Prof. Atul H. Karode2, Prof. S.R.Suralkar3
1

NMU University, SSBT College of Engineering and Technology, Department of E&TC, Jalgaon, Maharashtra, India
2

NMU University, SSBT College of Engineering and Technology, Department of E&TC, Jalgaon, Maharashtra, India

NMU University, SSBT College of Engineering and Technology, Department of E&TC, Jalgaon, Maharashtra, India

Abstract: In order to improve patient diagnosis various

image processing software are developed to extract useful information from medical images. Hematologist makes the microscopic study of human blood which led to a need of methods, including microscope colour imaging, segmentation, classification, and clustering that can allow the identification of patients suffering from leukemia. Leukemia is related with blast white blood cell (WBC). The nonspecific nature of the signs and symptoms of ALL often leads to wrong diagnosis so hematologist also find difficulty for blast cell classification hence manual classification of blood cells is time-consuming and susceptible to error. Therefore fast, accurate and automatic identification of different blood cells is required. This paper has proposed automatic Otsus threshold blood cell segmentation method along with image enhancement and arithmetic for WBC segmentation. kNN classifier has been utilized to classify blast cells from normal lymphocyte cells. The system is applied for 108 images available in public image dataset for the study of leukemia. This method gives 93% accuracy.

Keywords: Acute Lymphoblastic Leukemia (ALL), WBC segmentation, Public image dataset, FAB (FrenchAmerican-British) classification, kNN classifier.

1. INTRODUCTION
Leukemia is the blood cancer related with white blood cells [1]. It is a bone marrow disorder that arises when abnormal white blood cell begins to continuously replicate itself. These cells do not function normally which is to fight off infections. As they accumulate, they inhibit the production of other normal blood cells in the marrow, leading to anemia, bleeding, and recurrent infections. Over time, the leukemic cells spread through the bloodstream where they continue to divide, sometimes forming tumors and damaging organs such as the kidney and liver. Acute leukemia is classified according to the French-American- British (FAB) classification into two types: Acute Lymphoblastic Leukemia (ALL) and Acute Myelogenous Leukemia (AML). Acute leukemia is a rapidly progressing disease that affects mostly cells that are unformed (not yet fully developed or differentiated). ALL is most common in children while AML mainly affects adults but can occur in children and adolescents. In 2009, it is estimated that approximately 31,490 individuals will be diagnosed with leukemia and 44,510 individuals will die of the disease in the United States. Volume 2, Issue 3 May June 2013

The early and fast identification of the leukemia type, greatly aids in providing the appropriate treatment for the particular type. Its detection starts with a complete blood count (CBC) [2]. If the count is abnormal, the patient is suggested to perform bone marrow biopsy. Therefore, to confirm the presence of leukemic cells, a study of morphological bone marrow and peripheral blood slide analysis is done. In order to classify the abnormal cells in their particular types and subtype of leukemia, a hematologist will observe some cells under a light microscopy looking for the abnormalities presented in the nucleus or cytoplasm of the cells. The clinical behavior of the disease can be predicted using this classification and accordingly treatment should be given to the patient. In leukemia disease, large numbers of abnormal white blood cells are produced by bone marrow due to unknown cause. In pathology manual detection of leukemia is done which is time consuming as well as costly due to high cost pathology instruments. Hence automatic technique is adopted for fast and accurate results. In this technique image of blood sample is processed and nucleus part is segmented and finally cells are classified whether they are blast or normal one [19].

2. THE ACUTE LEUKEMIA DETECTION SYSTEM

Figure 1 shows the block diagram of a proposed system [4]. It consists of various functional modules. The main two steps in the proposed system are image segmentation and classification. The input image of blood slide is fed to the system. Blood has four main elements to ensure it fulfills its functions viz red blood cells, white blood cells, platelets, plasma [5].

Figure 1 Block diagram of a proposed system Page 147

International Journal of Emerging Trends & Technology in Computer Science (IJETTCS)

Web Site: www.ijettcs.org Email: editor@ijettcs.org, editorijettcs@gmail.com Volume 2, Issue 3, May June 2013 ISSN 2278-6856
White blood cell composition and concentration in the blood gives valuable information and plays a crucial role in the diagnosis of different diseases. Hence the very first module in the proposed system is white blood cell identification. White blood cells fall into five categories: Neutrophil, Eosinophil, Basophil, Monocyte and Lymphocyte [6]. The uncontrollable growth of lymphocyte is responsible for leukemia. Therefore lymphocyte cells are identified among white blood cells by segmentation. Then the features of lymphocytes such as area, perimeter, circularity, solidity etc are extracted. Then these features are applied to the classifier module where the classification among normal cells and blast cells occurs and detection of leukemia takes place. The most common method adopted for leukemia classification is the FAB method [3]; nowadays it has been updated with the immunologic classification which it is not image-based. Otherwise, the FAB method is still valid for image-based morphological classification. Various classifiers such as linear, kNN, fuzzy etc. are used for classification. desired colour vectors [10]. Jiang et al. presented a novel white blood cell segmentation method by combining two techniques, scale space filtering and watershed clustering [11]. In this scheme, two components of WBC, nucleus and cytoplasm, are extracted respectively using different methods. First, a sub image containing WBC is separated from the cell image. Then, scale space filtering is used to extract the nucleus region from the sub image. Later, watershed clustering in 3-D HSV (Hue, Saturation, Value) histogram is processed to extract cytoplasm region. Finally, morphological operations are performed to obtain the entire connective scheme successfully which avoids the variety and complexity in image space and can effectively extract various WBC regions from images of peripheral blood smear. The K-Mean clustering method is utilized by Sinha and Ramakrishnan to locate the white blood cell nuclei to identify the white blood cell on the image following by a crop operation to separate the entire cell from the image scene. However, the method of cropping the entire cell in order to get the real area of the whole cell is not clearly shown [12]. Theera-Umpon used a fuzzy C-Mean clustering to segment single cell images of white blood cells in the bone marrow into two regions, i.e., nucleus and non-nucleus. The computational time increases if the clusters number is greater than 2 [13].

3. IMAGE SEGMENTATION REVIEW

Segmentation means decomposition of image in different part for various applications [17]. The segmentation step is very crucial because the accuracy of the subsequent feature extraction and classification depends on the correct segmentation of white blood cells. It is also a difficult and challenging problem due to the complex nature of the cells and uncertainty in the microscopic image. Therefore, this step is the most important challenge in many literatures and improvement of cell segmentation has been the most common effort in many research works. Many researchers have given different methods for image segmentation such as threshold-based, edge-based, region-based or clustering methods, such as, fuzzy-C mean clustering and K-mean clustering. Cseke used automatic thresholding method (1979). Threshold techniques cannot always produce meaningful results since no spatial information is used during the selection of the segmentation threshold [7]. They are often combined with mathematical morphology operations. The technique was 92% accurate. Liao presents an accurate segmentation method for white blood cells [8]. A simple thresholding approach is applied to give initial labels to pixels in the blood cell images. The algorithm is based on priori information about blood smear images. Then the labels are adjusted with a shape detection method based on large regional context information to produce meaningful results. This scheme is only suitable if the shape of each white blood cell boundary is close to a circle. Edge detection method can also be meaningful for segmentation but it is applicable only when there is good contrast between foreground and background. (Piuri and Scotti) [9]. S. H. Rezatofighi et al. have proposed the segmentation of white blood cell nucleus based on GramSchmidt orthogonalization process for amplifying the Volume 2, Issue 3 May June 2013

4. IMAGE SEGMENTATION ALGORITHM

The proposed method for the segmentation of blood cell (leukocytes) is given below [14]. Step 1: Input the colour blood slide image to the system. Step 2: Convert the colour image into grayscale image. Step 3: Enhance contrast of the grayscale image by histogram equalization method (A). Step 4: To adjust image intensity level apply linear contrast stretching to gray scale image (B). Step 5: Obtain the image I1=B+A to brighten all other image components except cell nucleus. Step 6: Obtain the image I2=I1-A to highlight the entire image objects along with cell nucleus. Step 7: Obtain the image I3=I1+I2 to remove all other components of blood with minimum effect of distortion over nucleus. Step 8: To reduce noise, preserve edges and increase the darkness of the nuclei implement 3-by-3 minimum filter on the image I3. Step 9: Apply a global threshold Otsus method on image I3. Step 10: Using the threshold value in above step convert I3 to binary image. Step 11: To remove small pixel groups use morphological opening. Step 12: To form objects connect the neighboring pixels. Step 13: By applying the size test removal of all objects that are less than 50% of average RBC area is done. It is observed that this method of segmentation yields better results than that of previous methods. Page 148

International Journal of Emerging Trends & Technology in Computer Science (IJETTCS)

Web Site: www.ijettcs.org Email: editor@ijettcs.org, editorijettcs@gmail.com Volume 2, Issue 3, May June 2013 ISSN 2278-6856 5. FEATURE EXTRACTION
Feature extraction means to transfer the input data into different set of features. In image processing this is a technique of redefining a large set of redundant data into a set of features (or feature vector) of reduced dimension [15]. In this paper three features of lymphocyte cells have been observed viz, area, perimeter and circularity because shape of the nucleus is important feature for differentiation of blasts. All the features are extracted from the binary equivalent image of the nucleus with nonzero pixels representing the nucleus region. Area: The area was determined by counting the total number of none zero pixels within the image region. Perimeter: It was measured by calculating distance between successive boundary pixels. Circularity: This is a dimensionless parameter which changes with surface irregularities and is defined as, Circularity = 4* Pi* Area/ Perimerer2 Figure 2 Healthy blood images examples (a), blood with ALL blasts (b). (a1-2) and (b1-4) are zoomed subplots of the (a) and (b) images centered on lymphocytes and lymphoblast respectively.

8. EXPERIMENTAL RESULTS
The proposed technique has been applied on 108 peripheral blood smear images obtained from the public dataset as mentioned earlier. A microscopic blood image of size 2592 1944 is considered for evaluation [3]. As mentioned in section 4, algorithm applied to input image. The resulting images of segmentation are shown in figure 3, 4, 5 and 6.

6. CLASSIFICATION
Based on the features extracted in above step, classifier classifies the lymphocyte cells as blast or normal cells. Classification is the task of assigning to the unknown test vector, a label from one of the known classes [16]. The Knearest neighbor (kNN) decision rule has been a ubiquitous classification tool with good scalability. Past experience has shown that the optimal choice of K depends upon the data, making it laborious to tune the parameter for different applications. The k-NearestNeighbours (kNN) is a non-parametric method of classification. It is simple but very effective in many cases. Here also kNN has been utilized to classify blast cells from normal white blood cells.

Figure 3 Original image

7. DATABASE
For study purpose a public supervised image datasets (ALL-IDB) [18] is provided by Fabio Scotti to test and fairly compare algorithms for cell segmentation and classification of the ALL disease. There are two types of datasets are available. The ALL-IDB1 can be used both for testing segmentation capability of algorithms, as well as the classification systems and image preprocessing methods and ALL-IDB2 has segmented WBCs to test the classification of blast cells. The examples of ALLIDB1 dataset images are shown in figure 2. Figure 4 White blood cells with white spots over nucleus

Figure 5 Final segmented image In the result, numbers of WBCs are also counted. For a given figure 3, numbers of WBCs present are 11. WBCs are separated from other blood components by white spots over nuclei. In the final segmented image only WBCs are Volume 2, Issue 3 May June 2013 Page 149

International Journal of Emerging Trends & Technology in Computer Science (IJETTCS)

Web Site: www.ijettcs.org Email: editor@ijettcs.org, editorijettcs@gmail.com Volume 2, Issue 3, May June 2013 ISSN 2278-6856
kept with darker nucleus by removing all noisy components by minimum filter. After this segmentation lymphocyte cells are detected from all WBCs by blast segmentation and features of lymphocyte are calculated. Depending on those morphological features it is decided whether it is leukemic slide image or not. Final lymphocyte blast segmented image is shown in figure 6. [3] Ruggero Donida Labati IEEE Member, Vincenzo Piuri IEEE Fellow, Fabio Scotti IEEE Member, University of degli Studi di Milano, Department of Information Technologies, via Bramante 65, 26013 Crema, Italy, ALL-IDB: The Acute Lymphoblastic Leukemia Image Database For Image Processing, 2011 IEEE. [4] Wei-Liang Tai, Department of Biomedical Informatics, Asia University, Taiwan, Blood Cell Image Classification Based on Hierarchical SVM, 2011 IEEE. [5] Waidah Ismail, Department of Information Systems, Computing and Mathematics, Brunel University, Automatic Detection And Classification Of Leukemia Cells, June 2012. [6] Mostafa Mohamed, Behrouz Far, Department of Electrical and Computer Engineering, University of Calgary, Calgary, Canada, An Enhanced Threshold Based Technique for White Blood Cells Nuclei Automatic Segmentation, 14th International Conference on e-Health Networking, Applications and Services 2012. [7] Cseke, I., Geometry GIS System House Ltd. Budapest 1025, Felso Zoldmali 128-1 30. Hungary A fast segmentation scheme for white blood cell images, Proceedings of the 11th IAPR International Conference on Pattern Recognition Image, Speech and Signal Analysis, Aug. 30-Sept. 1992, Netherlands, pp: 530-533. [8] Qingmin Liao, Yingying Deng, Department of Electronic Engineering, Tsinghna University, P.R.China, An Accurate Segmentation Method for White Blood Cell Images, 0-7803-7584-x/OU$17.00 @ 2002 IEEE. [9] Vincenzo Piuri, Fabio Scotti University of Milan, Department of Information Technologies, via Bramante 65, 26013 Crema, Italy, Morphological Classification of Blood Leucocytes by Microscope Images, CIMSA 2004 -IEEE lntemational Conference on Computationvl Intelligence far Memrement Systems and Applications Boston, MA. USA, 14-16 July 2004. [10] S. H. Rezatofighi, Control and Intelligent Processing Center of Excellence, University of Tehran, Tehran, Iran, A New Approach to White Blood Cell Nucleus Segmentation Based on Gram-Schmidt Orthogonalization, International Conference on Digital Image Processing, IEEE 2009. [11] Jiang, K., Q.X. Jiang and Y. Xiong, Tsinghua University, Beijing, A novel white blood cell segmentation scheme using scale-space filtering and watershed clustering Mach. Learning Cybernetics, 5: 2820-2825, Proceedings of the Second International Conference on Machine Learning and Cybernetics, xi, 2-5 November 2003. [12] Neelam Sinha and A. G. Ramakrishnan, Department of Electrical Engineering Indian Institute of Science Page 150

Figure 6 Lymphocyte blast segmented image This method is applied on all 108 images. The accuracy of 93% is obtained using this method.

9. CONCLUSION
The main aim of this paper is blood slide image segmentation followed by feature extraction to detect leukemia. Shape features of nucleus such as area, perimeter, circularity etc. are considered for better accuracy of detection. Leukemia detection with proposed features were classified using kNN classifier giving overall accuracy of 93%. Furthermore the system should be robust to excessive staining and touching cells. Results obtained encourage future works which includes classification of lymphoblast into various subtypes. Alternate techniques can be investigated for stain independent blood smear image segmentation and leukemia type classification.

REFERENCES
[1] Catherine Haworth, Ad Heppleston, Ph Morris Jones, Rha Campbell, Dik Evans, Mk Palmer, From the Royal Manchester Children's Hospital, Pendlebury, Manchester M27 JHA, and the *Department of Statistics, Christie Hospital and Holt Radium Institute, Withington, Manchester M20 9BX, Routine bone inarrow examination in the management of acute lymphoblastic leukaemia of childhood, J Clin Pathol 1981 ;34:483-485. [2] Hayan T. Madhloom1, Sameem Abdul Kareem 2, Department of Artificial Intelligence, Faculty of Computer Science and Information, A Robust Feature Extraction and Selection Method for the Recognition of Lymphocytes versus Acute Lymphoblastic Leukemia, International Conference on Advanced Computer Science Applications and Technologies, 2012. Volume 2, Issue 3 May June 2013

International Journal of Emerging Trends & Technology in Computer Science (IJETTCS)

Web Site: www.ijettcs.org Email: editor@ijettcs.org, editorijettcs@gmail.com Volume 2, Issue 3, May June 2013 ISSN 2278-6856
Bangalore, Automation of Differential Blood Count, 2003 IEEE. [13] Nipon Theera-Umpon, Electrical Engineering Department, Chiang Mai University, Chiang Mai, Thailand, Breast Abnormality Detection in Mammograms Using Fuzzy Inference System, The International Conference on Fuzzy Systems 2005 IEEE. [14] H.T. Madhloom, S.A. Kareem, University of Malaya, Malaysia, An Automated White Blood Cell Nucleus Localization and Segmentation using Image Arithmetic and Automatic Threshold, Journal of Applied Sciences ,2010. [15] Subrajeet Mohapatra , Department of Electrical Engineering National Institute of Technology Rourkela Rourkela- 769008, Orissa, Automated Cell Nucleus Segmentation and Acute Leukemia Detection in Blood Microscopic Images, International Conference on Systems in Medicine and Biology 16-18 December 2010. [16] Yang Song, Department of Computer Science and Engineering, the Pennsylvania State University, USA, IKNN: Informative K-Nearest Neighbor Pattern Classification, Springer-Verlag Berlin Heidelberg 2007. [17] Digital Image Processing by Gonzalez Woods and Eddins. [18] http://www.dti.unimi.it/fscotti/all [19] Rohit Pathology, Dr. P.N.Patil, Jalgaon, Maharashtra, India.

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