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Automated method for the rapid and precise estimation of adherent cell culture characteristics from phase contrast microscopy images

Jaccard, N; Griffin, LD; Keser, A; Macown, RJ; Super, A; Veraitch, FS; Szita, N; (2014) Automated method for the rapid and precise estimation of adherent cell culture characteristics from phase contrast microscopy images. Biotechnology and Bioengineering , 111 (3) pp. 504-517. 10.1002/bit.25115. Green open access

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Abstract

The quantitative determination of key adherent cell culture characteristics such as confluency, morphology and cell density is necessary for the evaluation of experimental outcomes and to provide a suitable basis for the establishment of robust cell culture protocols. Automated processing of images acquired using phase contrast microscopy (PCM), an imaging modality widely used for the visual inspection of adherent cell cultures, could enable the non-invasive determination of these characteristics. We present an image-processing approach that accurately detects cellular objects in PCM images through a combination of local contrast thresholding and post-hoc correction of halo artifacts. The method was thoroughly validated using a variety of cell lines, microscope models and imaging conditions, demonstrating consistently high segmentation performance in all cases and very short processing times (< 1s per 1208 × 960 pixels image). Based on the high segmentation performance, it was possible to precisely determine culture confluency, cell density and the morphology of cellular objects, demonstrating the wide applicability of our algorithm for typical microscopy image processing pipelines. Furthermore, PCM image segmentation was used to facilitate the interpretation and analysis of fluorescence microscopy data, enabling the determination of temporal and spatial expression patterns of a fluorescent reporter. We created a software toolbox (PHANTAST) that bundles all the algorithms and provides an easy to use graphical user interface. Source-code for MATLAB and ImageJ is freely available under a permissive open-source license.

Type: Article
Title: Automated method for the rapid and precise estimation of adherent cell culture characteristics from phase contrast microscopy images
Open access status: An open access version is available from UCL Discovery
DOI: 10.1002/bit.25115
Publisher version: http://dx.doi.org/10.1002/bit.25115
Language: English
Additional information: This is an open access article under the terms of the Creative Commons Attribution License [http://creativecommons.org/licenses/by/3.0/], which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Keywords: adherent cells, cell density, confluency, image-processing, morphology, on-line monitoring, phase contrast microscopy
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Computer Science
URI: https://discovery.ucl.ac.uk/id/eprint/1408725
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