A call to action: democratizing single molecule fluorescence microscopy

Fluorescence-based optical techniques are developing rapidly, giving access to high spatiotemporal information on live biological systems with single molecule sensitivity. However, these techniques are typically restricted to expert labs and are not easilyaccessibleto the general user. While the development of customized systems and their wider distribution is difficult, as it requires expert manpower, software developments are easy to distribute. However, in reality only few users outside an expert community are exploring and using these tools. This is due to theusabilityof the software which often requires expert skills to operate and is neither intuitive nor easy to use. These issues of accessibility and usability limit the spread of state-of-the-art techniques. And while accessibility of custom instrumentation is difficult to solve, the accessibility and usability of software is an easier target. In this perspective, therefore, we concentrate on the software issue and examine the major translational barriers that prevent biologists from adopting the available fluorescence microscopy techniques. We discuss key developments in the field such as open-source tools, standardized file formats and AI-driven analysis platforms, and suggest a roadmap to bring advanced tools to a wider community.