Low-cost and open-source super-resolution fluorescence microscope with autofocus for teaching and research

Abstract

Super-resolution fluorescence microscopy revolutionized optical microscopy by breaking the theoretical resolution limit and thus providing an unprecedented view of biological structure in exquisite detail. Additionally, the relative ease of use and implementation for some super-resolution microscopes provided hope that this technology would be widely available and accessible. Despite initial excitement, super-resolution microscopy is still largely limited to facilities that can afford expensive commercial instruments or with personnel experienced in optical engineering that can construct an instrument. Recently, the open scientific hardware movement has attempted to make scientific instrumentation more widely available and accessible in a similar way to the open software movement. The widespread availability and affordability of prototyping devices, such as three-dimensional printers and interfaces (e.g., Arduino) have all contributed to unlocking the potential of the open hardware movement, turning expensive "black box" hardware into open-source, affordable, and robust instrumentation. Many others have recognized the potential of describing low-cost implementations of super-resolution microscope designs. However, we have found these microscope designs lacking in the documentation and details required to meet the requirements of open-source hardware. In this paper, we attempt to provide the documentation, details, and instructions necessary for the construction of an open-source and low-cost super-resolution microscope at a targeted cost of <$15,000.