Area-based Image Analysis Algorithm for Quantification of Macrophage-fibroblast Cocultures
| Author(s) | Borjigin, Tohn | |
| Author(s) | Boddupalli, Anuraag | |
| Author(s) | Sullivan, Millicent O. | |
| Date Accessioned | 2022-04-13T14:03:03Z | |
| Date Available | 2022-04-13T14:03:03Z | |
| Publication Date | 2022-02-15 | |
| Description | This article was originally published in Journal of Visualized Experiments. The version of record is available at: https://dx.doi.org/10.3791/63058. This article will be embargoed until 02/15/2024. | en_US |
| Abstract | Quantification of cells is necessary for a wide range of biological and biochemical studies. Conventional image analysis of cells typically employs either fluorescence detection approaches, such as immunofluorescent staining or transfection with fluorescent proteins or edge detection techniques, which are often error-prone due to noise and other non-idealities in the image background. We designed a new algorithm that could accurately count and distinguish macrophages and fibroblasts, cells of different phenotypes that often colocalize during tissue regeneration. MATLAB was used to implement the algorithm, which differentiated distinct cell types based on differences in height from the background. A primary algorithm was developed using an area-based method to account for variations in cell size/structure and high-density seeding conditions. Non-idealities in cell structures were accounted for with a secondary, iterative algorithm utilizing internal parameters such as cell coverage computed using experimental data for a given cell type. Finally, an analysis of coculture environments was carried out using an isolation algorithm in which various cell types were selectively excluded based on the evaluation of relative height differences within the image. This approach was found to accurately count cells within a 5% error margin for monocultured cells and within a 10% error margin for cocultured cells. | en_US |
| Sponsor | This work was funded in part by the National Institutes of Health (R01 AR067247) and in part by the Delaware INBRE program, supported by a grant from the National Institute of General Medical Sciences-NIGMS (P20 GM103446) from the National Institutes of Health and the State of Delaware. The contents of the manuscript do not necessarily reflect the views of the funding agencies. | en_US |
| Citation | Borjigin, T., Boddupalli, A., Sullivan, M. O. Area-based Image Analysis Algorithm for Quantification of Macrophage-fibroblast Cocultures. J. Vis. Exp. (180), e63058, doi:10.3791/63058 (2022). | en_US |
| ISSN | 1940-087X | |
| URL | https://udspace.udel.edu/handle/19716/30775 | |
| Language | en_US | en_US |
| Publisher | Journal of Visualized Experiments | en_US |
| Title | Area-based Image Analysis Algorithm for Quantification of Macrophage-fibroblast Cocultures | en_US |
| Type | Article | en_US |
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