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用于高通量测量遗传模式生物施加力的彩色聚二甲基硅氧烷微柱阵列。

Colored polydimethylsiloxane micropillar arrays for high throughput measurements of forces applied by genetic model organisms.

机构信息

Department of Physics, Indian Institute of Science , Bangalore 560012, India.

DBS-TIFR , Homi Bhabha Road, Mumbai 400005, India.

出版信息

Biomicrofluidics. 2015 Jan 29;9(1):014111. doi: 10.1063/1.4906905. eCollection 2015 Jan.

Abstract

Measuring forces applied by multi-cellular organisms is valuable in investigating biomechanics of their locomotion. Several technologies have been developed to measure such forces, for example, strain gauges, micro-machined sensors, and calibrated cantilevers. We introduce an innovative combination of techniques as a high throughput screening tool to assess forces applied by multiple genetic model organisms. First, we fabricated colored Polydimethylsiloxane (PDMS) micropillars where the color enhances contrast making it easier to detect and track pillar displacement driven by the organism. Second, we developed a semi-automated graphical user interface to analyze the images for pillar displacement, thus reducing the analysis time for each animal to minutes. The addition of color reduced the Young's modulus of PDMS. Therefore, the dye-PDMS composite was characterized using Yeoh's hyperelastic model and the pillars were calibrated using a silicon based force sensor. We used our device to measure forces exerted by wild type and mutant Caenorhabditis elegans moving on an agarose surface. Wild type C. elegans exert an average force of ∼1 μN on an individual pillar and a total average force of ∼7.68 μN. We show that the middle of C. elegans exerts more force than its extremities. We find that C. elegans mutants with defective body wall muscles apply significantly lower force on individual pillars, while mutants defective in sensing externally applied mechanical forces still apply the same average force per pillar compared to wild type animals. Average forces applied per pillar are independent of the length, diameter, or cuticle stiffness of the animal. We also used the device to measure, for the first time, forces applied by Drosophila melanogaster larvae. Peristaltic waves occurred at 0.4 Hz applying an average force of ∼1.58 μN on a single pillar. Our colored microfluidic device along with its displacement tracking software allows us to measure forces applied by multiple model organisms that crawl or slither to travel through their environment.

摘要

测量多细胞生物施加的力在研究其运动生物力学方面很有价值。已经开发了几种技术来测量这种力,例如应变计、微加工传感器和校准的悬臂梁。我们介绍了一种创新的技术组合,作为一种高通量筛选工具,用于评估多个遗传模式生物施加的力。首先,我们制造了彩色聚二甲基硅氧烷(PDMS)微柱,其颜色增强了对比度,使得更容易检测和跟踪由生物体驱动的微柱位移。其次,我们开发了一个半自动图形用户界面来分析微柱位移的图像,从而将每个动物的分析时间减少到几分钟。颜色的添加降低了 PDMS 的杨氏模量。因此,使用 Yeoh 超弹性模型对 PDMS 复合材料进行了表征,并使用基于硅的力传感器对微柱进行了校准。我们使用我们的设备来测量在琼脂表面上移动的野生型和突变型秀丽隐杆线虫施加的力。野生型秀丽隐杆线虫在单个微柱上施加的平均力约为 1μN,总平均力约为 7.68μN。我们表明,秀丽隐杆线虫的中部施加的力大于其两端。我们发现,身体壁肌肉有缺陷的秀丽隐杆线虫突变体在单个微柱上施加的力明显较低,而对外力感应有缺陷的突变体与野生型动物相比,每个微柱施加的平均力仍然相同。施加在每个微柱上的平均力与动物的长度、直径或表皮硬度无关。我们还首次使用该设备测量了黑腹果蝇幼虫施加的力。蠕动波以 0.4Hz 的频率发生,在单个微柱上施加的平均力约为 1.58μN。我们的彩色微流控设备及其位移跟踪软件允许我们测量通过其环境爬行或滑行的多个模型生物施加的力。

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