A programmable platform for probing cell migration and proliferation.

IF 6.6 3区 医学 Q1 ENGINEERING, BIOMEDICAL APL Bioengineering Pub Date : 2024-10-29 eCollection Date: 2024-12-01 DOI:10.1063/5.0209547
Jillian Cwycyshyn, Cooper Stansbury, Walter Meixner, James B Hoying, Lindsey A Muir, Indika Rajapakse
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Abstract

The advent of advanced robotic platforms and workflow automation tools has revolutionized the landscape of biological research, offering unprecedented levels of precision, reproducibility, and versatility in experimental design. In this work, we present an automated and modular workflow for exploring cell behavior in two-dimensional culture systems. By integrating the BioAssemblyBot® (BAB) robotic platform and the BioApps™ workflow automater with live-cell fluorescence microscopy, our workflow facilitates execution and analysis of in vitro migration and proliferation assays. Robotic assistance and automation allow for the precise and reproducible creation of highly customizable cell-free zones (CFZs), or wounds, in cell monolayers and "hands-free," schedulable integration with real-time monitoring systems for cellular dynamics. CFZs are designed as computer-aided design models and recreated in confluent cell layers by the BAB 3D-Bioprinting tool. The dynamics of migration and proliferation are evaluated in individual cells using live-cell fluorescence microscopy and an in-house pipeline for image processing and single-cell tracking. Our robotics-assisted approach outperforms manual scratch assays with enhanced reproducibility, adaptability, and precision. The incorporation of automation further facilitates increased flexibility in wound geometry and allows for many experimental conditions to be analyzed in parallel. Unlike traditional cell migration assays, our workflow offers an adjustable platform that can be tailored to a wide range of applications with high-throughput capability. The key features of this system, including its scalability, versatility, and the ability to maintain a high degree of experimental control, position it as a valuable tool for researchers across various disciplines.

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用于探测细胞迁移和增殖的可编程平台。
先进的机器人平台和工作流程自动化工具的出现彻底改变了生物研究的面貌,为实验设计提供了前所未有的精确性、可重复性和多功能性。在这项工作中,我们提出了一种在二维培养系统中探索细胞行为的自动化模块化工作流程。通过将 BioAssemblyBot® (BAB) 机器人平台和 BioApps™ 工作流程自动装置与活细胞荧光显微镜相结合,我们的工作流程促进了体外迁移和增殖实验的执行和分析。通过机器人辅助和自动化,可在细胞单层中精确、可重复地创建高度定制的无细胞区(CFZ)或伤口,并与细胞动态实时监测系统进行 "免提"、可调度的整合。无细胞区由计算机辅助设计模型设计,并通过 BAB 三维生物打印工具在汇合细胞层中重新创建。利用活细胞荧光显微镜以及内部图像处理和单细胞跟踪管道,对单个细胞的迁移和增殖动态进行评估。我们的机器人辅助方法在可重复性、适应性和精确性方面均优于人工划痕检测。自动化的加入进一步提高了伤口几何形状的灵活性,并允许在多种实验条件下进行并行分析。与传统的细胞迁移测定不同,我们的工作流程提供了一个可调整的平台,可根据广泛的应用进行定制,并具有高通量能力。该系统的主要特点,包括可扩展性、多功能性和保持高度实验控制的能力,使其成为各学科研究人员的重要工具。
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来源期刊
APL Bioengineering
APL Bioengineering ENGINEERING, BIOMEDICAL-
CiteScore
9.30
自引率
6.70%
发文量
39
审稿时长
19 weeks
期刊介绍: APL Bioengineering is devoted to research at the intersection of biology, physics, and engineering. The journal publishes high-impact manuscripts specific to the understanding and advancement of physics and engineering of biological systems. APL Bioengineering is the new home for the bioengineering and biomedical research communities. APL Bioengineering publishes original research articles, reviews, and perspectives. Topical coverage includes: -Biofabrication and Bioprinting -Biomedical Materials, Sensors, and Imaging -Engineered Living Systems -Cell and Tissue Engineering -Regenerative Medicine -Molecular, Cell, and Tissue Biomechanics -Systems Biology and Computational Biology
期刊最新文献
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