Critical comparison of image analysis workflows for quantitative cell morphological evaluation in assessing cell response to biomaterials
Critical comparison of image analysis workflows for quantitative cell morphological evaluation in assessing cell response to biomaterials
复制标题
用于定量细胞形态学评估的图像分析工作流程的批判性比较，以评估细胞对生物材料的反应
作者：
K. Ravikumar;Sven P. Voigt;S. Kalidindi;B. Basu
定量图像分析是通过研究细胞在大小、形状、数量和方向方面的形态学变化来理解细胞命运过程的重要工具。在这种情况下，这项工作系统地探讨了荧光显微镜图像的定量分析所涉及的主要挑战，并提出了一种新的协议，同时将其结果与广泛使用的ImageJ分析进行比较。值得一提的是，荧光显微镜是迄今为止最广泛用于植入式生物材料的生物相容性分析（观察细胞命运变化）。在这项研究中，我们采用了两种不同的图像分析工具集：（a）传统的ImageJ软件，（B）最近开发的自动化数字图像分析框架，称为ImageMKS。虽然ImageJ为图像分析提供了强大的工具集，但它需要复杂的用户专业知识来设计和迭代地优化分析工作流程。该工作流程主要包括一系列图像变换，通常涉及去噪和标记特征。另一方面，ImageMKS在很大程度上自动化了图像分析协议，从而减轻了用户偏见对最终结果的影响。这方面是解决使用的C2C12小鼠成肌细胞生长在聚（偏二氟乙烯）（PVDF）为基础的聚合物基板的案例研究。特别是，我们使用了一些荧光显微镜图像，这些小鼠成肌细胞生长在基于PVDF的纳米生物复合材料的电场的影响下。除了MKS工作流程由于其自动化而需要更少的用户时间之外，还观察到ImageMKS工作流程始终产生更可靠的结果，这些结果与先前报告的实验研究更好地相关。
Quantitative image analysis is an important tool in understanding cell fate processes through the study of cell morphological changes in terms of size, shape, number, and orientation. In this context, this work explores systematically the main challenges involved in the quantitative analysis of fluorescence microscopy images and also proposes a new protocol while comparing its outcome with the widely used ImageJ analysis. It is important to mention that fluorescence microscopy is by far most widely used in biocompatibility analysis (observing cell fate changes) of implantable biomaterials. In this study, we employed two different image analyses toolsets: (a) the conventionally employed ImageJ software, and (b) a recently developed automated digital image analyses framework, called ImageMKS. While ImageJ offers a powerful toolset for image analyses, it requires sophisticated user expertise to design and iteratively refine the analyses workflow. This workflow primarily comprises a sequence of image transformations that typically involve de-noising and labeling of features. On the other hand, ImageMKS automates the image analyses protocol to a large extent, and thereby mitigates the influence of the user bias on the final results. This aspect is addressed using a case study of C2C12 mouse myoblast cells grown on poly(vinylidene difluoride) (PVDF) based polymeric substrates. In particular, we used a number of fluorescence microscopy images of these mouse myoblasts grown on PVDF-based nanobiocomposites under the influence of electric field. In addition to the MKS workflows requiring much less user time because of their automation, it was observed that ImageMKS workflows consistently produced more reliable results that correlated better with the previously reported experimental studies.
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DOI：
10.2217/nnm.12.204
发表时间：
2013-11
期刊：
Nanomedicine (London, England)
影响因子：
--
作者：
Crowder SW;Liang Y;Rath R;Park AM;Maltais S;Pintauro PN;Hofmeister W;Lim CC;Wang X;Sung HJ
通讯作者：
Sung HJ
影响因子：
3.7
作者：
Serena E;Figallo E;Tandon N;Cannizzaro C;Gerecht S;Elvassore N;Vunjak-Novakovic G
通讯作者：
Vunjak-Novakovic G
影响因子：
19
作者：
Driscoll MK;Danuser G
通讯作者：
Danuser G
人类干细胞形态和分化的几何控制。
DOI：
10.1039/c0ib00016g
发表时间：
2010-08
期刊：
Integrative biology : quantitative biosciences from nano to macro
影响因子：
--
作者：
Wan LQ;Kang SM;Eng G;Grayson WL;Lu XL;Huo B;Gimble J;Guo XE;Mow VC;Vunjak-Novakovic G
通讯作者：
Vunjak-Novakovic G
影响因子：
2.7
作者：
Lamprecht, Michael R.;Sabatini, David M.;Carpenter, Anne E.
通讯作者：
Carpenter, Anne E.