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Scratch assay microscopy: A reaction–diffusion equation approach for common instruments and data
Mathematical Biosciences ( IF 4.3 ) Pub Date : 2020-10-02 , DOI: 10.1016/j.mbs.2020.108482
Alessio Gnerucci 1 , Paola Faraoni 2 , Elettra Sereni 2 , Francesco Ranaldi 2
Affiliation  

Scratch assay is an easy and widely used “in vitro” technique to study cell migration and proliferation. In this work we focus on its modelling and on the capability to distinguish between these two phenomena that the simpler and common models are not able to disentangle.

We adapted a model based on reaction–diffusion equation for being used with common microscopy instruments/data and therefore taking place in the gap between simpler modelling approaches and complex ones. An optimized image analysis pipeline and numerical least-squares fit provide estimates of the scratch proliferation and diffusion coefficients l and D.

This work is intended as a first of a series in which the model is tested and its robustness and reproducibility are evaluated. Test samples were NIH3T3 cells scratch assays with proliferation and migration stimulated by varying the foetal bovine serum amount in the culture medium (10%, 7.5%, 5% and 2.5%). Results demonstrate, notwithstanding an expected lD anticorrelation, the model capability to disentangle them. The 7.5% serum treatment can be identified as the model sensitivity limit. Treat–control l and D variations showed an intra-experiment reproducibility (±0.05h and ±200μm2h respectively) consistent with single fit typical uncertainties (±0.02h and ±300μm2h respectively).



中文翻译:

刮擦试验显微镜:用于常见仪器和数据的反应扩散方程方法

刮擦测定法是一种容易且广泛使用的“体外”技术,用于研究细胞迁移和增殖。在这项工作中,我们将重点放在其建模上以及区分这两种现象的能力,而这两种现象是较简单的通用模型无法解开的。

我们改编了基于反应扩散方程的模型,用于常见的显微镜仪器/数据,因此发生在简单建模方法与复杂建模方法之间的空白中。优化的图像分析管道和数值最小二乘拟合可提供划痕扩散和扩散系数的估计值d

这项工作旨在作为测试模型并评估其鲁棒性和再现性的系列文章中的第一篇。测试样品是NIH3T3细胞刮擦试验,通过改变培养基中胎牛血清的量(10%,7.5%,5%和2.5%)刺激增殖和迁移。结果证明,尽管有预期-d反相关,模型可以解开它们。血清7.5%的治疗可以确定为模型敏感性极限。治疗控制d 变化显示出实验内的可重复性(±005H±200μ2H 分别)与单一拟合典型不确定性(±002H±300μ2H 分别)。

更新日期:2020-10-04
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