Abstract
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.
Highlights
Live cell imaging is widely used in various bioscience experiments for a better understanding of dynamic cellular behaviors such as migration [1], division [2,3], differentiation [4], interaction with the environment [5], and organelle-level events [6]
This paper reports an incubator embedded cell culture imaging system, termed EmSight, which employs the Fourier ptychographic microscopy (FPM) for high resolution bright field imaging along with mid-resolution fluorescence imaging functionality
EmSight is a compact imaging system designed for live-cell imaging within standard biological incubators, usually within a volume of 200 liters
Summary
Live cell imaging is widely used in various bioscience experiments for a better understanding of dynamic cellular behaviors such as migration [1], division [2,3], differentiation [4], interaction with the environment [5], and organelle-level events [6]. In the most common way to perform live cell imaging, one builds a specialized incubation and imaging chamber onto a conventional microscope, and one images the cultured cells directly on the microscope stage [7] Recent advances with this method include robotic well plate transport systems [8] to increase throughput and the implementation of sufficiently compact and robust systems that can reside within an incubator [9]. The digital refocusing [20] capability associated with FPM processing, can allow postprocessing and correct defocus problems associated with cell culture imaging This is a marked benefit as the user can retain and analyze images that might be rejected with conventional imaging techniques. We summarize the results and discuss future directions and possible applications of the system
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