Question

does it possible to get difference between images in OpenCV of pixel in Matrix form

Answer 1

What do you mean by difference?
If you have two matrices with the same size and type, you can compute the difference:

cv::Mat::zeros m1(2,2,CV_32FC1);
cv::Mat::ones m2(2,2,CV32FC1);
std::cout << (m2-m1) << std::endl;