Damage to DNA and especially double strand breaks (DSBs) play a critical role both in the emergence

Damage to DNA and especially double strand breaks (DSBs) play a critical role both in the emergence and treatment of cancer. A single double strand break may be enough to bring about alterations in the genome that will eventually lead to cancer or cell death. γH2AX is a sensitive assay for measure DNA double strand breaks (DSBs) which are considered as most lethal damages to the DNA, leading to cell death. The method includes labeling the DSBs with fluorescent antibodies and counting these bright spots or foci, each representing one DSB. So by knowing the number of foci quantification of the DSBs is done, which is in response to DNA damaging agents. Radiobiologists count the number of DSBs in the nucleus manually, which appears as bright foci after staning with fluorescent tagged antibody.. So biologist counts the number of bright spots on the nucleus, which gives the amount of DSBs. Each layer of nucleus captured in terms of slices of images. After acquiring the slices of the nucleus, it needs to be merged together to manually count number of DSBs.

i = imread('particle.jpg');
I = rgb2gray(i);
BW3= edge(I,'roberts');
subplot (2,2,1);
imshow(I);
title('original');
subplot(2,2,2); 
imshow(BW1);
imshow(BW3); 
title('Roberts'); 
Read particle.jpg.
RGB = imread('particle.jpg');
imshow(RGB);
I = rgb2gray(RGB);
threshold = graythresh(I);
bw = im2bw(I,threshold);
imshow(bw)
bw = bwareaopen(bw,30);

se = strel('disk',2);
bw = imclose(bw,se);
bw = imfill(bw,'holes');

imshow(bw)
[B,L] = bwboundaries(bw,'noholes');

imshow(label2rgb(L, @jet, [.5 .5 .5]))
hold on
for k = 1:length(B)
boundary = B{k};
plot(boundary(:,2), boundary(:,1), 'w', 'LineWidth', 2)
end
metric = 4*pi*area/perimeter^2.
stats = regionprops(L,'Area','Centroid');

threshold = 0.94;

for k = 1:length(B)

boundary = B{k};

delta_sq = diff(boundary).^2;
perimeter = sum(sqrt(sum(delta_sq,2)));

area = stats(k).Area;

metric = 4*pi*area/perimeter^2;

metric_string = sprintf('%2.2f',metric);

if metric > threshold
centroid = stats(k).Centroid;
plot(centroid(1),centroid(2),'ko');
end

text(boundary(1,2)-35,boundary(1,1)+13,metric_string,'Color','y',...
'FontSize',14,'FontWeight','bold');

end

title(['Metrics closer to 1 indicate that ',...
'the object is approximately round']);

[Walter Roberson]

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