function out_mask = WBC_SegMadhloom(input_image, show_images) %This function is built to segment the nuclei of white blood cells % the input

is an image and an option to show the resluts on screen or not (1 or 0) %and the number of white blood cells % For more information abou the algorithm please refere to: % "An Efficient Technique for White Blood Cells Nuclei Automatic Segmentation", % IEEE SMC conference 2012 % The algorithm in this function is an implementation if the paper: % H. T. Madhloom, S. A. Kareem, H. Ariffin, A. A. Zaidan, H. O. Alanazi and % B. B. Zaidan, "An Automated White Blood Cell Nucleus Localization and % Segmentation using Image Arithmetic and Automatic Threshold," Journal of % Applied Sciences, vol. 10, no. 11, pp. 959-966, 2010. if(ischar(input_image)==0) a1 = input_image; input_image = 'Memory'; else a1=imread(input_image); end; if show_images == 1, figure;imshow(a1);title('Input image'); end %% Convert to gray scale image a=rgb2gray(a1); if show_images == 1, figure;imshow(a);title('Input image in gray scale'); end; %% Adjust image intensity values with a linear contrast stretching L=imadjust(a); if show_images == 1, figure;imshow(L);title('L: Input image with intensity values adjusted by a linear contrast stretching'); end; %% Enhance contrast using histogram equalization H = histeq(a); if show_images == 1, figure;imshow(H);title('H: Enhance contrast using histogram equalization'); end %% Brighten most of the details in the image except the nucleus R1=imadd(L,H); if show_images == 1, figure;imshow(R1);title('R1: Brighten most of the details in the image except the nucleus'); end %% Highlight all the objects and its borders in the image including the cell nucleus R2 = imsubtract(R1,H); if show_images == 1, figure;imshow(R2);title('R2: Highlight all the objects and its borders in the image including the cell nucleus'); end %% Remove almost all the other blood components while retaining the nucleus with minimum affect of distortion on the nucleus part of the white blood cell.

R3=imadd(R1,R2); if show_images == 1, figure;imshow(R3);title('R3: Remove almost all the other blood components while retaining the nucleus with minimum affect of distortion on the nucleus part of the white blood cell.'); end %check histogram % figure; % hi=imhist(R3); % hi1=hi(1:2:256); % horz=1:2:256; % bar(horz,hi1); % % %axis([0 255 0 1400]); % % %set(gca, 'xtick', 0:50:255); % % %set(gca, 'ytick', 0:2000:15000); % xlabel('Gray level' ); % ylabel('No of pixels' ); % title('Histogram before opening the image'); % 'Time elapsed to calculate image preparations (algorithm)' % toc(programStart) filterStart=tic; %===================== %implements a 3-by-3 minimum filter %reduce noise, preserve edges and increase the darkness of the nuclei for i=1:1 R3=ordfilt2(R3,1,ones(3,3)); end % 'Time elapsed to calculate 3-by-3 minimum filter' % toc(filterStart) %figure; imshow(R3); %check histogram afteraplying the minimum filter % figure; % hi=imhist(R3); % hi1=hi(1:2:256); % horz=1:2:256; % bar(horz,hi1); % % %axis([0 255 0 10000]); % % %set(gca, 'xtick', 0:50:255); % % %set(gca, 'ytick', 0:2000:15000); % xlabel('Gray level' ); % ylabel('No of pixels' ); % title('Histogram before opening the image'); %===================================================== %% Global threshold using Otsus method level=graythresh(R3); bw=im2bw(R3,level); %Complement image

bw = imcomplement(bw); if show_images == 1, figure;imshow(bw);title('Global threshold using Otsus method '); end %% Show the results out_mask = bw; if show_images == 1 %Re-count objects in the image cells = bwconncomp(bw); no_of_WBCs=cells.NumObjects; a2 = a1; for j=1:no_of_WBCs %return the coordinates for the pixels in object j [r, c] = find(bwlabel(bw)==j); rc = [r c]; %marking the location of the nuclei of white blood cells by a white %colour for i=1:max(size(rc)) a2(rc(i,1),rc(i,2),:)=uint8(a1(rc(i,1),rc(i,2),:)*1.5); end end h = figure(10); set(h, 'Name', 'WBC segmentation results','NumberTitle','off', 'OuterPosition',[1 1 1600 600]) subplot(1,3,1);imshow(a1); [~, name ext] = fileparts(input_image); title(['The original image [' name ext ']']); subplot(1,3,2);imshow(a2); title('The original image with the white spot(s) over the WBC nucleus(ei)'); %show the final segmented image bw1 = imcomplement(bw); amask = a; amask(bw1) = 255; subplot(1,3,3);imshow(amask); title('The final segmented image mask'); % disp(['Total program time is: ' num2str(toc(programStart))]); % % % % % % Check Histogram of masked gray scale image figure(5); % imshow(a(bw)) hi=imhist(amask); % imshow(amask) hi1=hi(1:2:256); horz=1:2:256; bar(horz,hi1);

% % % % % % %

axis([0 255 0 1400]); % %set(gca, 'xtick', 0:50:255); % %set(gca, 'ytick', 0:2000:15000); xlabel('Gray level' ); ylabel('No of pixels' ); Save images path = fileparts(input_image); imwrite(a1, [path '\a1Mah.bmp'], 'bmp') imwrite(a2, [path '\a2Mah.bmp'], 'bmp') imwrite(bw1, [path '\abwMah.bmp'], 'bmp')

end