Aparanji, S and Kamat, S (2022) A simple flow-cytometry based image processing algorithm for analysing cell-death mechanisms in radiotherapy. In: Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XX 2022, 20-24 February 2022, Virtual, Online at San Francisco, California, United States,.
Full text not available from this repository.Abstract
In-vitro testing of novel photodynamic therapy/radiotherapy procedures relies heavily on the use of different assays to fully probe various parameters such as cytotoxicity or cell-death pathways. These assays utilise sometimes expensive dyes or antibodies, along with cumbersome sample preparation for flow-cytometry. In this work, we propose a novel image-processing algorithm that uses the flow cytometry plots obtained through a Propidium Iodide based live-dead assay on cancerous and non-cancerous cells to deduce the possible cell-death mechanisms in the process of radiotherapy. Propidium Iodide (PI) is a membrane-impermeable dye taken up by those cells with loss of cell membrane integrity, and does not give any information about the integrity of intracellular components or cellular death pathways. In our novel image-processing algorithm, we determine the centroid of the Forward Scatter (FSC) and the Side Scatter (SSC) cytometer plots of such a PI assay, after suitable clustering. This algorithm is initially applied to an unirradiated control cell population where the FSC centroid gives an estimate of the mean cell size, while the SSC centroid gives the baseline granularity of the cell population. Thereafter, the centroids of the FSC and the SSC plots are calculated for the irradiated cell population, and the deviation in these centroids calculated. These differences are correlated to change in average cell size and denaturation/granularity, and serve as a useful substitute for the cell death mechanism. This can potentially pave the way for in-situ qualitative cell-death analysis in large-volume in-vitro studies in a cost-effective manner. © COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only.
| Item Type: | Conference Paper |
|---|---|
| Publication: | Progress in Biomedical Optics and Imaging - Proceedings of SPIE |
| Publisher: | SPIE |
| Additional Information: | The copyright for this article belongs to the SPIE. |
| Keywords: | Cell culture; Cell proliferation; Clustering algorithms; Cost effectiveness; Fiber lasers; Flow cytometry; Photodynamic therapy; Radiotherapy; Tumors, Cell death mechanism; Cell populations; Cell-size; Dead assays; Image processing algorithm; In-vitro testing; Propidium iodide; Propidium iodide live-dead assay; Raman fiber lasers; Simple++, Cell death |
| Department/Centre: | Division of Biological Sciences > Biochemistry Division of Interdisciplinary Sciences > Centre for Nano Science and Engineering |
| Date Deposited: | 22 Jun 2022 09:51 |
| Last Modified: | 22 Jun 2022 09:51 |
| URI: | https://eprints.iisc.ac.in/id/eprint/73944 |
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