用双光子聚合技术制备了一种用于拉伸载荷下单细胞-细胞连接成像的多材料平台

IF 3 4区 医学 Q3 ENGINEERING, BIOMEDICAL
Jordan Rosenbohm, Grayson Minnick, Bahareh Tajvidi Safa, Amir Monemian Esfahani, Xiaowei Jin, Haiwei Zhai, Nickolay V. Lavrik, Ruiguo Yang
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引用次数: 2

摘要

我们之前报道了一个由IP-S光树脂与双光子聚合(TPP)制成的单细胞粘附微拉伸测试仪(SCAμTT),用于研究单细胞-细胞连接在规定拉伸载荷下的力学。该平台的一个主要限制是IP-S的自身荧光,这是用于TPP制造的光树脂,它会显著增加背景信号,使拉伸细胞的荧光成像变得困难。在这项研究中,我们报告了一种新的SCAμTT平台的设计和制造,该平台可以减轻自身荧光,并证明了其在单细胞对的相互连接被拉伸时成像的能力。通过采用IP-S和IP-Visio(一种减少自身荧光的光树脂)的双材料设计,我们发现平台的自身荧光显著减少。此外,通过将孔径与金涂层集成到衬底上,几乎完全减轻了自身荧光对成像的影响。利用这个新平台,我们展示了对一对上皮细胞进行成像的能力,当它们被拉伸到250%的应变时,我们可以观察到连接处的破裂和f -肌动蛋白的收缩,同时记录连接处超过800kpa的应力积累。这里提出的平台和方法可以潜在地详细研究细胞-细胞连接的力学和机械转导,并改进机械生物学应用中其他TPP平台的设计。图形抽象
本文章由计算机程序翻译,如有差异,请以英文原文为准。

A multi-material platform for imaging of single cell-cell junctions under tensile load fabricated with two-photon polymerization

A multi-material platform for imaging of single cell-cell junctions under tensile load fabricated with two-photon polymerization

We previously reported a single-cell adhesion micro tensile tester (SCAμTT) fabricated from IP-S photoresin with two-photon polymerization (TPP) for investigating the mechanics of a single cell-cell junction under defined tensile loading. A major limitation of the platform is the autofluorescence of IP-S, the photoresin for TPP fabrication, which significantly increases background signal and makes fluorescent imaging of stretched cells difficult. In this study, we report the design and fabrication of a new SCAμTT platform that mitigates autofluorescence and demonstrate its capability in imaging a single cell pair as its mutual junction is stretched. By employing a two-material design using IP-S and IP-Visio, a photoresin with reduced autofluorescence, we show a significant reduction in autofluorescence of the platform. Further, by integrating apertures onto the substrate with a gold coating, the influence of autofluorescence on imaging is almost completely mitigated. With this new platform, we demonstrate the ability to image a pair of epithelial cells as they are stretched up to 250% strain, allowing us to observe junction rupture and F-actin retraction while simultaneously recording the accumulation of over 800 kPa of stress in the junction. The platform and methodology presented here can potentially enable detailed investigation of the mechanics of and mechanotransduction in cell-cell junctions and improve the design of other TPP platforms in mechanobiology applications.

Graphical abstract

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来源期刊
Biomedical Microdevices
Biomedical Microdevices 工程技术-工程:生物医学
CiteScore
6.90
自引率
3.60%
发文量
32
审稿时长
6 months
期刊介绍: Biomedical Microdevices: BioMEMS and Biomedical Nanotechnology is an interdisciplinary periodical devoted to all aspects of research in the medical diagnostic and therapeutic applications of Micro-Electro-Mechanical Systems (BioMEMS) and nanotechnology for medicine and biology. General subjects of interest include the design, characterization, testing, modeling and clinical validation of microfabricated systems, and their integration on-chip and in larger functional units. The specific interests of the Journal include systems for neural stimulation and recording, bioseparation technologies such as nanofilters and electrophoretic equipment, miniaturized analytic and DNA identification systems, biosensors, and micro/nanotechnologies for cell and tissue research, tissue engineering, cell transplantation, and the controlled release of drugs and biological molecules. Contributions reporting on fundamental and applied investigations of the material science, biochemistry, and physics of biomedical microdevices and nanotechnology are encouraged. A non-exhaustive list of fields of interest includes: nanoparticle synthesis, characterization, and validation of therapeutic or imaging efficacy in animal models; biocompatibility; biochemical modification of microfabricated devices, with reference to non-specific protein adsorption, and the active immobilization and patterning of proteins on micro/nanofabricated surfaces; the dynamics of fluids in micro-and-nano-fabricated channels; the electromechanical and structural response of micro/nanofabricated systems; the interactions of microdevices with cells and tissues, including biocompatibility and biodegradation studies; variations in the characteristics of the systems as a function of the micro/nanofabrication parameters.
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