Joint Department of Biomedical Engineering, University of North Carolina-Chapel Hill/North Carolina State University, Raleigh 27695, North Carolina, United States.
Comparative Medicine Institute, North Carolina State University, Raleigh 27695, North Carolina, United States.
ACS Sens. 2021 Mar 26;6(3):985-994. doi: 10.1021/acssensors.0c02172. Epub 2021 Mar 3.
The ability to measure microtissue contraction in vitro can provide important information when modeling cardiac, cardiovascular, respiratory, digestive, dermal, and skeletal tissues. However, measuring tissue contraction in vitro often requires the use of high number of cells per tissue construct along with time-consuming microscopy and image analysis. Here, we present an inexpensive, versatile, high-throughput platform to measure microtissue contraction in a 96-well plate configuration using one-step batch imaging. More specifically, optical fiber microprobes are embedded in microtissues, and contraction is measured as a function of the deflection of optical signals emitted from the end of the fibers. Signals can be measured from all the filled wells on the plate simultaneously using a digital camera. An algorithm uses pixel-based image analysis and computer vision techniques for the accurate multiwell quantification of positional changes in the optical microprobes caused by the contraction of the microtissues. Microtissue constructs containing 20,000-100,000 human ventricular cardiac fibroblasts (NHCF-V) in 6 mg/mL collagen type I showed contractile displacements ranging from 20-200 μm. This highly sensitive and versatile platform can be used for the high-throughput screening of microtissues in disease modeling, drug screening for therapeutics, physiology research, and safety pharmacology.
体外测量微组织收缩的能力在心脏、心血管、呼吸、消化、皮肤和骨骼组织建模时可以提供重要信息。然而，体外测量组织收缩通常需要每个组织构建物使用大量细胞，并结合耗时的显微镜和图像分析。在这里，我们提出了一种廉价、通用、高通量的平台，可使用一步批量成像在 96 孔板配置中测量微组织收缩。更具体地说，光纤微探针嵌入微组织中，收缩程度通过光纤末端发出的光信号的偏折来测量。可以使用数字相机同时测量板上所有填充孔的信号。一种算法使用基于像素的图像分析和计算机视觉技术，对微组织收缩引起的光学微探针的位置变化进行准确的多孔定量分析。含有 20,000-100,000 个人心室心肌成纤维细胞 (NHCF-V) 的 6mg/mL Ⅰ型胶原微组织构建物显示出 20-200μm 的收缩位移。该高灵敏度、通用的平台可用于疾病建模、治疗药物筛选、生理学研究和安全药理学中的微组织高通量筛选。
