Recent advances in optical imaging with single molecules beyond the diffraction limit (e.g., PALM, FPALM, STORM) have introduced a new requirement for fluorescent labels: fluorophores must be actively controlled (usually via photoswitching or photo-activation) to ensure that only a single emitter is switched on at a time in a diffraction-limited region. The location of each of these sparse molecules is precisely determined, and a super-resolution image is obtained from the summation of many successive rounds of photoactivation. The ultimate spatial resolution is determined by a number of factors, most importantly the total number of photons detected from each individual molecule.
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