A process for maintaining 3 dimensional orientation between a
tissue specimen and images of the area of investigation, to register histopathologic diagnoses of multiple locations within the specimen with corresponding locations on the surface of said area of investigation, by marking at least two fiduciary lines on the area of investigation; acquiring a fiduciary image of the tissue with the fiduciary lines; excising the tissue to form a
tissue specimen; inserting at least two parallel needles through said specimen; acquiring a specimen image of the specimen with inserted needles over an alignment grid; fixing the specimen by immersing the specimen; acquiring a fixed image of the
fixed specimen with the inserted needles over the alignment grid; forming a paraffin mold containing the
fixed specimen and inserted needles; injecting different
colored inks through the needles while withdrawing them from the
fixed specimen, so that different
colored needle tracks are formed in the specimen; sectioning the specimen to create specimen blocks having different
colored needle tracks; further sectioning the specimen to
cut the specimen blocks into specimen slices having different colored ink dots corresponding to the different colored needle tracks; forming
pathology images from the specimen slices; performing
histopathology analyses on the
pathology images; annotating the
pathology images with
histopathology annotations; aligning the annotations with the fixed image using the colored ink dots; determining shrinkage between the fixed image and the annotations by using the grid to compare the distance between the needles in the fixed image with the distance between the ink dots on the specimen slices; registering the fixed image to the specimen image to account for shrinkage caused by fixation, using locations of the needles in both of the images as landmarks; registering the specimen image to the fiduciary image to account for tissue translation and
soft tissue movement using the fiduciary lines and geographical features of said area of investigation as landmarks; registering the fiduciary image to the
reference image to account for tissue translation and
soft tissue movement using said geographical features; whereby annotations of histopathologic diagnoses are provided for multiple locations on or under the surface of the specimen that are registered to images of the specimen.