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. 2011 Jun 14;3(6):770–793. doi: 10.3390/v3060770

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© 2011 by the authors; licensee MDPI, Basel, Switzerland.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).

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Figure 5. — Super-resolution microscopy. Super-resolution microscopy obtains fluorescence-based spatial information with sub-diffraction (<200 nm) resolution. This is accomplished by using photo-activatable (PALM/fPALM) or photo-switchable (STORM) fluorophores, such that a sparse, spatially-separated sub-population of fluorophores is active at any one time. Upon photo-activation of the sub-population, the positions of the well-separated fluorophores can be determined with high spatial resolution by mapping the center of the diffraction-limited fluorescence signal. The activated fluorophores are then de-activated either by photobleaching (PALM/fPALM) or photo-switching, and a different sub-population is activated and localized. By repeating this process thousands of times, a composite, super-resolution image can be constructed using the mapped position of every fluorophore.