The initial form of SMLM systems was not only expensive but Super-resolution image is dependent on the performance of localization algorithms and the photophysical switching properties of the The resolution or quality of the reconstructed Therefore, SMLM is largely a computational imaging technique, built upon a simple configuration of a wide-field fluorescence microscope. The fundamental principle of SMLM is based on singlemolecule localization, in which a small subset of labeled fluorophores is sequentially turned “on,” then the centers of individualĮmitters are determined by localization algorithms at a nanometer precision, and the final reconstructed image is obtained afterĪccumulating localized positions from tens of thousands of imageįrames. Understand the biological systems at the molecular scale. These advantages, SMLM has quickly become an essential tool to To ∼5 nm7 can be obtained with standard illumination power density, much better than STED (∼50 nm) and SIM (∼100 nm). Photon counts (tens of thousands of photons per emitter throughīright emitters and long exposure time), a spatial resolution downĪPL Photon. Require complex and expensive optics for patterned illuminationĪnd, thus, is the most cost-effective approach to achieve superresolved imaging capability. Distinct from other super-resolution imaging techniques such as stimulated emission depletion (STED) or structured illumination microscopy (SIM), single-molecule localization Microscopy have revolutionized biological imaging by overcoming the fundamental diffraction barrier, recognized by a Nobel In the last decade, the advances in super-resolution fluorescence However, due to theĭiffraction-limited spatial resolution of optical microscopy, conventional fluorescence microscopy cannot visualize biological structures smaller than the diffraction-limited resolution (∼200 nm). Specificity and multiplex imaging capability. In the past few decades, fluorescence microscopy has significantly expanded our ability to study biological processes at theĬellular and subcellular level, on the strength of its molecular All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license Recent technical advances and their implications in serving the community of biomedical research. 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. Of optical microscopy has rapidly gotten out of the ivory tower and entered a new phase to address various challenging biomedical questions. 5, 060902 (2020) doi: 10.1063/5.0011731īiomedical Optical Imaging Laboratory, Departments of Medicine and Bioengineering, University of Pittsburgh,Īuthor to whom correspondence should be addressed: nearly 15 years since its initial debut, super-resolution localization microscopy that surpasses the diffraction-limited resolution barrier High throughput, high quality, and low costĬite as: APL Photon. Super-resolution localization microscopy: Toward Structured illumination microscopy using digital micro-mirror device and coherent lightĪpplied Physics Letters 116, 233702 (2020) įar-field unlabeled super-resolution imaging with superoscillatory illumination This paper was selected as an Editor’s Pickįluorescence polarization filtering for accurate single molecule localization Toward high throughput, high quality, andĬite as: APL Photonics 5, 060902 (2020) Super-resolution localization microscopy:
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