We have two different microscopic image of samples, based on intensity of gray in image1 i want to identify region in image 2 which has different intensity other then gray intensity in image1?
Or other question is can i get the difference of two image and if yes how to get image3 which is difference of image1 and image2?
Hi Ravi,
If I understand you correctly, you want to subtract image1 from image2, so you can see the "difference". If so, you have to just use the intensity subtraction, and subtract matrix intensities in the image 1 from the intensities in the image2. You can do this in the MATLAB really easy. I have attached an example here.
Beyond the question: do you want to see the objects in the image2 clearly? maybe you want to count the number of them? if so you have to use the segmentation algorithm, in which the background (like image1) can be calculated from the image2. The foreground(objects) can be separated.
One of classical segmentation algorithm is Ostu Method: https://en.wikipedia.org/wiki/Otsu%27s_method.
A MATLAB implementation can be found here: https://www.mathworks.com/help/images/ref/graythresh.html
Best,
JW
In Fiji this could be performed by Process -> Image Calculator -> select two images and select an operation (substract in your case)
According to your images you can achieve the following manipulation using MATLAB.
I1 = imread('Image1.tif');
I2 = imread('Image2.tif');
[rowsI1, colsI1]=size(I1);
I2=imresize(I2,[rowsI1, colsI1]);
O1 = imabsdiff(I1,I2) ;
figure ; imshow(O1)
O2 = 255-O1 ;
figure ; imshow(O2)
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