Graduate School of Engineering, Muroran Institute of Technology, 27-1 Mizumoto-cho, Muroran, Hokkaido, 050-8585, Japan.
Sci Rep. 2022 Oct 7;12(1):16884. doi: 10.1038/s41598-022-20598-6.
When studying physical cellular response observed by light microscopy, variations in cell behavior are difficult to quantitatively measure and are often only discussed on a subjective level. Hence, cell properties are described qualitatively based on a researcher's impressions. In this study, we aim to define a comprehensive approach to estimate the physical cell activity based on migration and morphology based on statistical analysis of a cell population within a predefined field of view and timespan. We present quantitative measurements of the influence of drugs such as cytochalasin D and taxol on human neuroblastoma, SH-SY5Y cell populations. Both chemicals are well known to interact with the cytoskeleton and affect the cell morphology and motility. Being able to compute the physical properties of each cell for a given observation time, requires precise localization of each cell even when in an adhesive state, where cells are not visually differentiable. Also, the risk of confusion through contaminants is desired to be minimized. In relation to the cell detection process, we have developed a customized encoder-decoder based deep learning cell detection and tracking procedure. Further, we discuss the accuracy of our approach to quantify cell activity and its viability in regard to the cell detection accuracy.
当通过光学显微镜研究观察到的物理细胞反应时,细胞行为的变化很难进行定量测量,通常只能在主观层面上进行讨论。因此,细胞特性是根据研究人员的印象进行定性描述的。在这项研究中,我们旨在定义一种全面的方法,根据在预设的视野和时间段内对细胞群体的统计分析,基于迁移和形态来估计细胞的物理活性。我们提出了对细胞的物理性质的定量测量,这些细胞是人类神经母细胞瘤 SH-SY5Y 细胞群体,这些化学物质都已知与细胞骨架相互作用,并影响细胞形态和运动性。能够计算给定观察时间内每个细胞的物理性质,即使在细胞处于黏附状态(细胞在视觉上不可区分)时,也需要对每个细胞进行精确的定位。此外,还希望尽量减少因污染物而导致的混淆风险。关于细胞检测过程,我们开发了一种基于定制的编码器-解码器的深度学习细胞检测和跟踪程序。此外,我们还讨论了我们的方法在量化细胞活性及其在细胞检测精度方面的可行性方面的准确性。