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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 - Biotechnol. Bioeng. (2013)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemical Engineering, University College London, Torrington Place, London, WC1E 7JE, United Kingdom; Centre for Mathematics and Physics in the Life Sciences and Experimental Biology, University College London, London, United Kingdom.

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Cell density estimation of mESC cultures based on PCM images. (A) Relationship (adjusted r2 = 0.89) between cell density (as measured after detachment) and culture confluency (determined using PHANTAST). (B) Changes in mean cell area during a typical mESC culture. Cells were counted using a live nuclear stain (Hoechst 33342). The insert shows the change in distance between nuclei during the same culture. (C) The relationship between cell density (as determined after detachment) and packing-corrected confluency (PCC), computed from the confluency and the distance to nearest nucleus-like feature using PHANTAST.
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fig05: Cell density estimation of mESC cultures based on PCM images. (A) Relationship (adjusted r2 = 0.89) between cell density (as measured after detachment) and culture confluency (determined using PHANTAST). (B) Changes in mean cell area during a typical mESC culture. Cells were counted using a live nuclear stain (Hoechst 33342). The insert shows the change in distance between nuclei during the same culture. (C) The relationship between cell density (as determined after detachment) and packing-corrected confluency (PCC), computed from the confluency and the distance to nearest nucleus-like feature using PHANTAST.

Mentions: For mESC, and colony-forming cell lines in general, cell density as determined using sacrificial counting methods is not linearly proportional to culture confluency ( Fig. 5A). Indeed, high variations in the area of individual cells were measured during the course of a culture ( Fig. 5B), consistent with typical pluripotent stem cells growth behavior (Harb et al., 2008). In general, confluency is therefore a poor predictor of cell density.


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 - Biotechnol. Bioeng. (2013)

Cell density estimation of mESC cultures based on PCM images. (A) Relationship (adjusted r2 = 0.89) between cell density (as measured after detachment) and culture confluency (determined using PHANTAST). (B) Changes in mean cell area during a typical mESC culture. Cells were counted using a live nuclear stain (Hoechst 33342). The insert shows the change in distance between nuclei during the same culture. (C) The relationship between cell density (as determined after detachment) and packing-corrected confluency (PCC), computed from the confluency and the distance to nearest nucleus-like feature using PHANTAST.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4260842&req=5

fig05: Cell density estimation of mESC cultures based on PCM images. (A) Relationship (adjusted r2 = 0.89) between cell density (as measured after detachment) and culture confluency (determined using PHANTAST). (B) Changes in mean cell area during a typical mESC culture. Cells were counted using a live nuclear stain (Hoechst 33342). The insert shows the change in distance between nuclei during the same culture. (C) The relationship between cell density (as determined after detachment) and packing-corrected confluency (PCC), computed from the confluency and the distance to nearest nucleus-like feature using PHANTAST.
Mentions: For mESC, and colony-forming cell lines in general, cell density as determined using sacrificial counting methods is not linearly proportional to culture confluency ( Fig. 5A). Indeed, high variations in the area of individual cells were measured during the course of a culture ( Fig. 5B), consistent with typical pluripotent stem cells growth behavior (Harb et al., 2008). In general, confluency is therefore a poor predictor of cell density.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemical Engineering, University College London, Torrington Place, London, WC1E 7JE, United Kingdom; Centre for Mathematics and Physics in the Life Sciences and Experimental Biology, University College London, London, United Kingdom.

Show MeSH
Related in: MedlinePlus