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BioMed Research International
Volume 2018, Article ID 7082154, 15 pages
Research Article

Colour Vignetting Correction for Microscopy Image Mosaics Used for Quantitative Analyses

1Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy
2Advanced Research Center on Electronic Systems for Information and Communication Technologies “E. De Castro” (ARCES), University of Bologna, Bologna, Italy
3Department of Computer Science and Engineering (DISI), University of Bologna, Bologna, Italy

Correspondence should be addressed to Alessandro Bevilacqua; ti.obinu@auqcaliveb.ordnassela

Received 21 February 2018; Revised 30 April 2018; Accepted 10 May 2018; Published 7 June 2018

Academic Editor: Jiang Du

Copyright © 2018 Filippo Piccinini and Alessandro Bevilacqua. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Image mosaicing permits achieving one high-resolution image, extending the visible area of the sample while keeping the same resolution. However, intensity inhomogeneity of the stitched images can alter measurements and the right perception of the original sample. The problem can be solved by flat-field correcting the images through the vignetting function. Vignetting correction has been widely addressed for grey-level images, but not for colour ones. In this work, a practical solution for the colour vignetting correction in microscopy, also facing the problem of saturated pixels, is described. In order to assess the quality of the proposed approach, five different tonal correction approaches were quantitatively compared using state-of-the-art metrics and seven pairs of partially overlapping images of seven different samples. The results obtained proved that the proposed approach allows obtaining high quality colour flat-field corrected images and seamless mosaics without employing any blending adjustment. In order to give the opportunity to easily obtain seamless mosaics ready for quantitative analysis, the described vignetting correction method has been implemented in an upgraded release of MicroMos (version 3.0), an open-source software specifically designed to automatically obtain mosaics of partially overlapped images.