Vignetting and photo-bleaching correction in automated fluorescence microscopy from an array of overlapping images

We propose a novel acquisition scheme and non-parametric multi-image based method for correcting illumination in fluorescence images. Our approach measures changes in intensity of the subject by moving the microscope stage at regularly spaced intervals, and exploits this information to learn the cor...

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Bibliographic Details
Published in2013 IEEE 10th International Symposium on Biomedical Imaging pp. 464 - 467
Main Authors Piccinini, Filippo, Bevilacqua, Alessandro, Smith, Kevin, Horvath, Peter
Format Conference Proceeding
LanguageEnglish
Published IEEE 01.04.2013
Subjects
Additives
Arrays
automated microscopy
Calibration
illumination correction
image processing
Lighting
Microscopy
mosaicing
Optical microscopy
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Summary:We propose a novel acquisition scheme and non-parametric multi-image based method for correcting illumination in fluorescence images. Our approach measures changes in intensity of the subject by moving the microscope stage at regularly spaced intervals, and exploits this information to learn the correction function. The acquisition process and learning are performed prior to imaging, and take only a few minutes. Afterwards, images can be corrected for vignetting and photobleaching effects on the fly. Our approach can be implemented in any microscope with a motorized stage, and does not require a reference calibration slide. Experiments demonstrate that our method outperforms standard approaches to illumination correction.
ISBN:1467364568
9781467364560
ISSN:1945-7928
DOI:10.1109/ISBI.2013.6556512