(D-146) Creating an Accessible Arduino-Powered Imaging Rig for Versatile Live Image Acquisition

The imaging rig designed in this study was composed of several components, including a PC with the open-source imaging software μManager, a microscope, a software-compatible digital eyepiece camera, an LED gooseneck illuminator, and an Arduino (Figure 1). Notably, the Arduino simplifies the process of loading new code onto the board, as it does not require additional hardware such as a programmer. A standard USB cable suffices for this purpose. 

The Arduino was programmed using an easily adjustable code in C++ as a shutter device and was then attached at digital Pin13 to a relay that controls an illumination device. To complete the imaging system, the left microscope eyepiece was replaced with a camera which was also connected to the PC and configured through μManager’s acquisition settings. The imaging software settings allowed for the customization of parameters to suit a range of experiments through the ‘Multi-S Asq.’ tab. 

In this specific application, the Arduino was set up to receive input from μManager via USB and trigger an output, which turned on the illuminator during image acquisition for one second every ten minutes for in vitro experiments. To protect the light-sensitive samples, a 3D-printed chamber cover was placed around the base of the microscope. 

Developmental assays were performed for each cell line by first preparing and then plating 107 cells on an agar plate. Once the system was configured within μManager, images were automatically acquired every ten minutes over a twenty-four hour period.

After successfully capturing and saving the data over the twenty-four hour experiments, the images were conveniently analyzed directly in ImageJ, fully integrated through μManager. Critical differences in both the timing and morphology of the developmental programs between the cell lines were made easily observable (Figure 2). Through comparison of the precisely captured timed images, ME3- knockout cells displayed a delayed developmental timing with smaller slug structures in comparison to the wildtype phenotype. PikG knockout cells show no development at all throughout the twenty-four hours. The phenotypic differences between the cell strains could be further quantified through ImageJ. 

The combination of software and hardware in the imaging setup enabled precise and automated image acquisition, avoiding prolonged illumination of the Dictyostelium cells, making an efficient and reliable system for performing long time course experiments. Beyond the current study, the extensive imaging rig can be successfully applied to various other experiments, such as chimeric assays, complementation experiments, and investigations of phagocytic and autophagic activity at low magnifications. Although there are similar technologies available at significantly higher costs, opting to design custom rigs comes with unparalleled creative license and autonomy at more accessible price points. The adaptability of this tool is vast, as it can be customized to meet specific demands of distinct experiments, providing limitless possibilities for observation, investigation, and analysis. 

Moreover, as this project strives to establish a more affordable and flexible version of existing technology, it thereby reduces the entry barriers for biomedical research studies worldwide but especially in developing nations. In resource-limited settings, these cost-effective and accessible live imaging microscope units can serve as versatile tools, potentially playing a crucial role in swiftly assessing, monitoring, and addressing various concerns, including infectious diseases. This study highlights the pivotal role of technology in enhancing scientific research by promoting accessibility and creative design, especially in the context of global health and development.  

This work was supported by the Gregory S. Call Student Research Fund and NSF. A special thanks to Dr. Marc Edwards for his unwavering trust and invaluable guidance throughout the research process.  

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