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The handheld digital microscope features a 3D-printed chassis to house its hardware, firmware, and rechargeable Li-ion battery with built-in power management. It incorporates an internal stainless-steel cage system to enclose and provide mechanical rigidity for the optics and imaging sensor. To reduce the microscope’s size, yet retain high spatial resolution, engineers devised an optical light path that uniquely folds back on itself using high reflectivity mirrors, thus significantly reducing internal volume. Imaging control and acquisition is performed using a secure web-based graphical user interface accessible via any wireless enabled device. The microscope serves as its own wireless access point thus obviating the need for a pre-existing network. This web interface enables multiple simultaneous connections and facilitates data sharing with clinicians, scientists, or other personnel as needed. Acquired images can be stored locally on the microscope server or on a removable SD card. Data can be securely downloaded to other devices using a range of industry standard protocols. Although the handheld digital microscope was originally developed for in-flight medical diagnosis in microgravity applications, prototypes were thoroughly ground-tested in a variety of environments to verify the accurate resolve of microbial samples for identification and compo-sitional analysis for terrestrial field use. Owing to its portability, other applications demanding rapid results may include research, education, veterinarian, military, contagion disaster response, telemedicine, and point-of-care medicine.