Single molecule localization microscopy

Filrstenberg A, Heilemann M (2013) Single-molecule localization microscopy— near-molecular spatial resolution in light microscopy with photoswitchable fluorophores. Phys Chem ChemPhys 15(36) 14919-14930... 

Single Molecule Localization Microscopy to Study Neuronal Microtubule Organization... 

Different fluorescence microscopy techniques are commonly used to explore the organization of the microtubule cytoskeleton. The resolution of these techniques is, however, limited by diffraction to approximately 250 nm, which makes them not suitable for nanoscale mapping of microtubule properties. Superresolution microscopy techniques that rely on single molecule localization (Single Molecule Localization Microscopy SMLM) combine high protein specificity, multi-color imaging, and a resolution in the order of 5-50 nm, making it an ideal tool to study the neuronal cytoskeleton and its properties. 

X. Qu et ah. Nanometer-localized multiple single-molecule fluorescence microscopy. Proc. Natl. Acad. Sci. USA 101(31), 11298-11303 (2004)... 

Herbert S, Soares H, Zimmer C et al (2012) Single-molecule localization super-resolution microscopy deeper and faster. Microsc Microanal 18 1419-1429... 

However, finding out the position of an object with arbitrary precision is not the same as resolution, which is about separating similar objects at small distances. Localization per se cannot provide superresolution. This is also why, although it had been known and used for decades [109,110] and even routinely applied to single molecules [125,126], localization alone has not provided nanoscale images. (Note that in spite the use of localization in the 1980 s and earlier, near-field optical microscopy still seemed to be the only way to attain nanoscale resolution up to the early 1990 s.) Resolution clearly requires a criterion to discern objects or molecules, the simplest of which is bright vs. dark. ... 

Manley, S., Gillette, J. M., Patterson, G. H., Shroff, H., Hess, H. E, Betzig, E., and Lippincott-Schwartz, J. 2008. High-density mapping of single-molecule trajectories with photoactivated localization microscopy, Nat Methods 5,155-157. 

---