Abstract
After nearly 15 years since its initial debut, super-resolution localization microscopy that surpasses the diffraction-limited resolution barrier of optical microscopy has rapidly gotten out of the ivory tower and entered a new phase to address various challenging biomedical questions. Recent advances in this technology greatly increased the imaging throughput, improved the imaging quality, simplified the sample preparation, and reduced the system cost, making this technology suitable for routine biomedical research. We will provide our perspective on the recent technical advances and their implications in serving the community of biomedical research.
Highlights
In the past few decades, fluorescence microscopy has significantly expanded our ability to study biological processes at the cellular and subcellular level, on the strength of its molecular specificity and multiplex imaging capability
In combination with high photon counts, a spatial resolution down to ∼5 nm7 can be obtained with standard illumination power density, much better than stimulated emission depletion (STED) (∼50 nm) and structured illumination microscopy (SIM) (∼100 nm)
The automated workflow enables the maintenance of consistent imaging conditions throughout the entire imaging process, which is crucial for generating reproducible image data
Summary
In the past few decades, fluorescence microscopy has significantly expanded our ability to study biological processes at the cellular and subcellular level, on the strength of its molecular specificity and multiplex imaging capability. The advances in super-resolution fluorescence microscopy have revolutionized biological imaging by overcoming the fundamental diffraction barrier, recognized by a Nobel Prize in 2014. In combination with high photon counts (tens of thousands of photons per emitter through bright emitters and long exposure time), a spatial resolution down to ∼5 nm can be obtained with standard illumination power density, much better than STED (∼50 nm) and SIM (∼100 nm). With these advantages, SMLM has quickly become an essential tool to understand the biological systems at the molecular scale. Scitation.org/journal/app latest advances in the technical development of SMLM systems for super-resolution microscopy and their implications in the field of biomedical research
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