I have 4 16 bit images that I took by rotating a sample 0, 90, 180, and 270 degrees respectively. Since I rotated the sample manually the fields of view in the images don’t line up perfectly. I’ve been trying align them in order to then make a composite image. I’ve tried Big Warp, SIFT align, and Align_RGB_planes so far with no luck. Also a side note, I’ve rotated all the images back to “0” degrees before applying these methods. I don’t want to end up with a stack of images, I only want one 2D image at the end if that helps. My images are of mouse skin (collagen is visible) with second harmonic generation microscopy if that’s important information.