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Automated stitching of microscope images of fluorescence in cells with minimal overlap

Title
Automated stitching of microscope images of fluorescence in cells with minimal overlap
Authors
Seo J.-H.Yang S.Kang M.-S.Her N.-G.Nam D.-H.Choi J.-H.Kim M.H.
Ewha Authors
김명희최장환
SCOPUS Author ID
김명희scopus; 최장환scopus
Issue Date
2019
Journal Title
Micron
ISSN
0968-4328JCR Link
Citation
Micron vol. 126
Keywords
Brain tumor cellFluorescence microscope imagesHigh-content screeningImage stitchingTile scan
Publisher
Elsevier Ltd
Indexed
SCI; SCIE; SCOPUS scopus
Document Type
Article
Abstract
The morphology of tumor cells is highly related to their phenotype and activity. To verify the drug response of a brain tumor patient, fluorescence microscope images of drug-treated patient-derived cells in each well are analyzed. Due to the limitation of the field of view (FOV), a large number of small FOVs are acquired to compose one complete microscope well. Here, we propose an automated method for accurately stitching tile-scanned fluorescence microscope images, even with noise and a narrow overlapping region between adjacent fields. The proposed method is based on intensity-based normalized cross-correlation (NCC) and a triangular method-based threshold. The proposed method's quantitative accuracy and the sensitivity of the input was compared to other existing stitching tools, MIST and FijiIS, setting manually stitched images as the ground truth. The test images were 20 samples of 3 × 3 grid images in three versions of the fluorescence channel. The distance between the location of each field and number of cells was determined for different input field overlap ranges (1%, 3%, 5%, and 10%), while the actual value was about 1.15%. The proposed method had a distance error of 1.5 pixels at an input overlap of 1%, showing the lowest minimum error at all channels. Regarding the difference in cell numbers, although the number of overlapping cells was always small because of the narrow overlapping range, the proposed method was able to generate the resultant image with the smallest difference. In addition, to confirm the size limitation of the proposed algorithm, the accuracy of stitching images of grid structures 3 × 3, 5 × 5, 10 × 10–20 × 20 was tested, showing consistent results. In conclusion, quantitative evaluation of the performance of the method proved its improved accuracy compared to other current state-of-art techniques, and it showed robust performance even with noise and a narrow overlapping region between adjacent fields. © 2019 Elsevier Ltd
DOI
10.1016/j.micron.2019.102718
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엘텍공과대학 > 컴퓨터공학과 > Journal papers
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