Super-resolution microscopy at its sharpest
What’s next then?
First, it is becoming important to explore how such high levels of precision can be reliably translated to biomedical imaging settings while coping with the complexity inherent to the biological world. For this reason, optics-driven advancements in super-resolution microscopy must involve close collaboration with biologists, to understand their needs and what questions need answers. Meaningful development is often initiated, and then driven, by asking the right questions and finding the right problem to solve, aligning efforts with real needs. Collaboration with (bio)chemists is also important to make sure labelling workflows are feasible and reliable.
Second, as spatial resolution approaches the size of fluorophores themselves, we enter what is often referred to as nanoscopy. Although this opens up exciting possibilities for studying proteins and potentially their conformational changes with unprecedented detail, it also comes with challenges. Labelling samples at such an extreme scale is difficult, as existing methods struggle to attach fluorescent tags just a few nanometres apart within cells. We also need to deal with the different photochemistry of two fluorophores spaced by less than 10 nm, as their photo-switching behaviour can be altered, reducing the probability of accurate localization. Importantly, this challenge is shared across multiple techniques, showing the importance of finding solutions.
Finally, the ability to image live cells over long periods of time is highly valued when it comes to studying dynamical processes in real time. However, super-resolution microscopy works best with fixed samples. Live cells are mobile, thus demanding careful optimization of the trade-off between spatial resolution, imaging speed and field of view. Hours-long imaging also raises challenges of phototoxicity and photobleaching, and it highlights the need for new fluorescent probes that are both stable and non-invasive over long periods.
Overall, this Focus issue attempts to capture a selection of recent developments and spark discussions about new opportunities and outstanding challenges to increase our understanding of the biological world. Through synergistic efforts from optics, biology and biochemistry, super-resolution microscopy is poised to keep developing, gain wider adoption and provide solutions to real-world biomedical challenges that can tangibly improve healthcare and clinical outcomes.
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