使用微芯片增强光片显微镜对细胞球体进行大规模高通量3D培养、成像和分析。
Large-scale high-throughput 3D culture, imaging, and analysis of cell spheroids using microchip-enhanced light-sheet microscopy.
作者信息
Zhu Tingting, Nie Jun, Yu Tingting, Zhu Dan, Huang Yanyi, Chen Zaozao, Gu Zhongze, Tang Jiang, Li Dongyu, Fei Peng
机构信息
School of Optical and Electronic Information - Wuhan National Laboratory for Optoelectronics - Advanced Biomedical Imaging Facility, Huazhong University of Science and Technology, Wuhan 430074, China.
Institute for Cell Analysis, Shenzhen Bay Laboratory, Shenzhen 518132, China.
出版信息
Biomed Opt Express. 2023 Mar 27;14(4):1659-1669. doi: 10.1364/BOE.485217. eCollection 2023 Apr 1.
Light sheet microscopy combined with a microchip is an emerging tool in biomedical research that notably improves efficiency. However, microchip-enhanced light-sheet microscopy is limited by noticeable aberrations induced by the complex refractive indices in the chip. Herein, we report a droplet microchip that is specifically engineered to be capable of large-scale culture of 3D spheroids (over 600 samples per chip) and has a polymer index matched to water (difference <1%). When combined with a lab-built open-top light-sheet microscope, this microchip-enhanced microscopy technique allows 3D time-lapse imaging of the cultivated spheroids with ∼2.5-µm single-cell resolution and a high throughput of ∼120 spheroids per minute. This technique was validated by a comparative study on the proliferation and apoptosis rates of hundreds of spheroids with or without treatment with the apoptosis-inducing drug Staurosporine.
结合微芯片的光片显微镜技术是生物医学研究中一种新兴的工具，显著提高了效率。然而，微芯片增强光片显微镜受到芯片中复杂折射率引起的明显像差的限制。在此，我们报道了一种液滴微芯片，该芯片经过特殊设计，能够大规模培养三维球体（每个芯片超过600个样本），并且其聚合物折射率与水匹配（差异<1%）。当与实验室构建的开放式光片显微镜结合使用时，这种微芯片增强显微镜技术能够以约2.5微米的单细胞分辨率对培养的球体进行三维延时成像，通量高达每分钟约120个球体。通过对数百个球体在有或没有用凋亡诱导药物星形孢菌素处理情况下的增殖和凋亡率进行比较研究，验证了该技术。
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