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Incubator embedded cell culture imaging system (EmSight) based on Fourier ptychographic microscopy

机译:基于傅立叶色谱分析的孵化器嵌入式细胞培养成像系统(EmSight)

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摘要

Multi-day tracking of cells in culture systems can provide valuable information in bioscience experiments. We report the development of a cell culture imaging system, named EmSight, which incorporates multiple compact Fourier ptychographic microscopes with a standard multiwell imaging plate. The system is housed in an incubator and presently incorporates six microscopes. By using the same low magnification objective lenses as the objective and the tube lens, the EmSight is configured as a 1:1 imaging system that, providing large field-of-view (FOV) imaging onto a low-cost CMOS imaging sensor. The EmSight improves the image resolution by capturing a series of images of the sample at varying illumination angles; the instrument reconstructs a higher-resolution image by using the iterative Fourier ptychographic algorithm. In addition to providing high-resolution brightfield and phase imaging, the EmSight is also capable of fluorescence imaging at the native resolution of the objectives. We characterized the system using a phase Siemens star target, and show four-fold improved coherent resolution (synthetic NA of 0.42) and a depth of field of 0.2 mm. To conduct live, long-term dopaminergic neuron imaging, we cultured ventral midbrain from mice driving eGFP from the tyrosine hydroxylase promoter. The EmSight system tracks movements of dopaminergic neurons over a 21 day period.
机译:在培养系统中对细胞进行多日跟踪可以为生物科学实验提供有价值的信息。我们报告了名为EmSight的细胞培养成像系统的开发,该系统结合了多个紧凑型傅里叶分型显微镜和标准多孔成像板。该系统位于培养箱中,目前装有六个显微镜。通过使用与物镜和管状镜相同的低倍率物镜,EmSight被配置为1:1成像系统,可在低成本CMOS成像传感器上提供大视场(FOV)成像。 EmSight通过以不同的照明角度捕获样品的一系列图像来提高图像分辨率。该仪器通过使用迭代傅立叶指纹图谱算法重建高分辨率图像。除了提供高分辨率的明场和相位成像,EmSight还能够以物镜的自然分辨率进行荧光成像。我们使用相位西门子星靶对系统进行了表征,并显示出相干分辨率提高了四倍(合成NA为0.42),景深为0.2 mm。为了进行实时的,长期的多巴胺能神经元成像,我们从酪氨酸羟化酶启动子驱动eGFP的小鼠中培养了腹侧中脑。 EmSight系统在21天的时间内跟踪多巴胺能神经元的运动。

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