The first evaluation was conducted on 30 sets of triple-staining prostate cancer samples, including H&E slides and IHC slides with two kinds of antigens, CK18 and HMCK, and we compare the proposed method with four image registration techniques for biological microscopic image alignment, including bUnwarpJ7, LeastSquares8, Elastic9 and CwR10. This evidence concerns the gene KRT18 and prostate carcinoma.