Abstract
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.
Highlights
IntroductionIt has been used for more than a century; the first attempts of panoramic photographs are found at war photography (e.g., during the American Civil War in 1860) [1]
Mosaicing has a key role for this purpose, since high-resolution images representing a whole sample are a valuable resource for pathologists and biologists in general [4]
We proposed a practical colour vignetting correction for mosaicing applications
Summary
It has been used for more than a century; the first attempts of panoramic photographs are found at war photography (e.g., during the American Civil War in 1860) [1]. Nowadays, they are widely used in oncology, in particular in histopathology [2]. As far as microscopy is concerned, various high-magnification microscopes are used in order to observe the fine details of biological specimens. They all suffer from a limited field of view (FOV) [3]. Errors propagated via geometric and photometric misalignments result in undesirable seams and object discontinuities that can be seen at the borders of the images
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