Research during the past 2 decades has showcased the
power of single-molecule
localization microscopy (SMLM) as a tool for exploring the nanoworld.
However, SMLM systems are typically available in specialized laboratories
and imaging facilities, owing to their expensiveness as well as complex
assembly and alignment procedure. Here, we lay out the blueprint of
a sturdy, rail-based, cost-efficient, multicolor SMLM setup that is
easy to construct and align in service of simplifying the accessibility
of SMLM. We characterize the optical properties of the design and
assess its capabilities, robustness, and stability. The performance
of the system is assayed using super-resolution imaging of biological
samples. We believe that this design will make SMLM more affordable
and broaden its availability.