A computer vision approach for analyzing label free leukocyte trafficking dynamics on a microvascular mimetic.
A computer vision approach for analyzing label free leukocyte trafficking dynamics on a microvascular mimetic.
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DOI：
10.3389/fimmu.2023.1140395
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发表时间：
2023
影响因子：
7.3
通讯作者：
McGrath JL
中科院分区：
文献类型：
--
作者：
Ahmad SD;Cetin M;Waugh RE;McGrath JL
高内涵成像技术与体外微生理系统（MPS）相结合，可以对由于使用人类细胞系而具有高度转化相关性的生理现象进行新颖的探索。过去，采用超薄纳米多孔氮化硅膜 (μSiM) 的 MPS 已被用来促进高放大倍数相差显微镜记录模拟人体血管微环境中的白细胞运输事件。值得注意的是，成像平面可以直接设置在 µSiM 设备的内皮界面上，从而高分辨率捕获内皮细胞 (EC) 和白细胞共培养物对不同刺激条件的反应。在该界面记录观察产生的大量数据可用于阐明与血管屏障功能障碍相关的疾病机制，例如脓毒症。这些记录中白细胞的出现是动态的，其特征、位置和时间都在变化。因此，传统的图像处理技术无法提取响应疾病状态的大量白细胞的时空分布和大量统计数据，需要劳动密集型的手动处理，这是该方法的一个重大限制。在这里，我们描述了一个机器学习管道，它使用语义分割算法和分类脚本，结合起来能够在共培养模拟物中进行自动化和无标签的白细胞运输分析。在表征白细胞时空行为时，开发的计算工具集与手动制表数据集具有明显的同等性，计算效率高，并且能够以半自动方式管理大型成像数据集。
High-content imaging techniques in conjunction with in vitro microphysiological systems (MPS) allow for novel explorations of physiological phenomena with a high degree of translational relevance due to the usage of human cell lines. MPS featuring ultrathin and nanoporous silicon nitride membranes (µSiM) have been utilized in the past to facilitate high magnification phase contrast microscopy recordings of leukocyte trafficking events in a living mimetic of the human vascular microenvironment. Notably, the imaging plane can be set directly at the endothelial interface in a µSiM device, resulting in a high-resolution capture of an endothelial cell (EC) and leukocyte coculture reacting to different stimulatory conditions. The abundance of data generated from recording observations at this interface can be used to elucidate disease mechanisms related to vascular barrier dysfunction, such as sepsis. The appearance of leukocytes in these recordings is dynamic, changing in character, location and time. Consequently, conventional image processing techniques are incapable of extracting the spatiotemporal profiles and bulk statistics of numerous leukocytes responding to a disease state, necessitating labor-intensive manual processing, a significant limitation of this approach. Here we describe a machine learning pipeline that uses a semantic segmentation algorithm and classification script that, in combination, is capable of automated and label-free leukocyte trafficking analysis in a coculture mimetic. The developed computational toolset has demonstrable parity with manually tabulated datasets when characterizing leukocyte spatiotemporal behavior, is computationally efficient and capable of managing large imaging datasets in a semi-automated manner.
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DOI：
10.1007/978-1-61779-207-6_18
发表时间：
2011-01-01
期刊：
CELL MIGRATION: DEVELOPMENTAL METHODS AND PROTOCOLS, SECOND EDITION
影响因子：
--
作者：
Elks, Philip M.;Loynes, Catherine A.;Renshaw, Stephen A.
通讯作者：
Renshaw, Stephen A.
影响因子：
3.1
作者：
Kaufmann, Ines;Hoelzl, Alwin;Thiel, Manfred
通讯作者：
Thiel, Manfred
影响因子：
5.8
作者：
Arganda-Carreras, Ignacio;Kaynig, Verena;Seung, H. Sebastian
通讯作者：
Seung, H. Sebastian
DOI：
10.1109/tit.1962.1057692
发表时间：
1962-01-01
期刊：
IRE TRANSACTIONS ON INFORMATION THEORY
影响因子：
--
作者：
HU, M
通讯作者：
HU, M
影响因子：
7.7
作者：
Joseph A;Chu CJ;Feng G;Dholakia K;Schallek J
通讯作者：
Schallek J