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Issue 95, 2014
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A rapid and effective vignetting correction for quantitative microscopy

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Abstract

Images acquired using optical microscopes are inherently subject to vignetting effects due to imperfect illumination and image acquisition. However, such vignetting effects hamper accurate extraction of quantitative information from biological images, leading to less effective image segmentation and increased noise in the measurements. Here, we describe a rapid and effective method for vignetting correction, which generates an estimate for a correction function from the background fluorescence without the need to acquire additional calibration images. We validate the usefulness of this algorithm using artificially distorted images as a gold standard for assessing the accuracy of the applied correction and then demonstrate that this correction method enables the reliable detection of biologically relevant variation in cell populations. A simple user interface called FlattifY was developed and integrated into the image analysis platform YeastQuant to facilitate easy application of vignetting correction to a wide range of images.

Graphical abstract: A rapid and effective vignetting correction for quantitative microscopy

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Publication details

The article was received on 04 Aug 2014, accepted on 14 Oct 2014 and first published on 14 Oct 2014


Article type: Paper
DOI: 10.1039/C4RA08110B
Author version available: Download Author version (PDF)
Citation: RSC Adv., 2014,4, 52727-52733
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    A rapid and effective vignetting correction for quantitative microscopy

    S. S. Lee, S. Pelet, M. Peter and R. Dechant, RSC Adv., 2014, 4, 52727
    DOI: 10.1039/C4RA08110B

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