In many clinical studies, including those of cancer, it is highly desirable to acquire images of whole tumour sections whilst retaining a microscopic resolution. A usual approach to this is to create a composite image by appropriately overlapping individual images acquired at high magnification under a microscope. A mosaic of these images can be accurately formed by applying image registration, overlap removal and blending techniques. We describe an optimised, automated, fast and reliable method for both image joining and blending. These algorithms can be applied to most types of light microscopy imaging. Examples from histology, from in vivo vascular imaging and from fluorescence applications are shown, both in 2D and 3D. The algorithms are robust to the varying image overlap of a manually moved stage, though examples of composite images acquired both with manually-driven and computer-controlled stages are presented. The overlap-removal algorithm is based on the cross-correlation method; this is used to determine and select the best correlation point between any new image and the previous composite image. A complementary image blending algorithm, based on a gradient method, is used to eliminate sharp intensity changes at the image joins, thus gradually blending one image onto the adjacent 'composite'. The details of the algorithm to overcome both intensity discrepancies and geometric misalignments between the stitched images will be presented and illustrated with several examples.

Date Published: 24 March 2005
PDF: 10 pages
Proc. SPIE 5701, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII, (24 March 2005); doi: 10.1117/12.590536

Published in SPIE Proceedings Vol. 5701:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII
Jose-Angel Conchello; Carol J. Cogswell; Tony Wilson, Editor(s)