Stochastic optical reconstruction microscopy

Rust, M. J., Bates, M. and Zhuang, X. (2006). Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM). Nat. Methods 3, 793-5. 

In another superresolution approach. Rust et al. have utilized controlled photoswitching of small molecule emitters for superresolution demonstrations [134] (STORM, for Stochastic Optical Reconstruction Microscopy). This method uses a Cy3-Cy5 emitter pair in close proximity in the presence of thiols that show a novel property restoration of Cy5 s photobleached emission can be achieved by brief pumping of the Cy3 molecule. In this way, the emission from a single Cy5 is turned on and off, again and again, to measure its position accurately multiple times. In the Moerner laboratory, a covalently bound Cy3-Cy5 dimer has been prepared as shown in Fig. 2.12C [135]. This molecule has the advantage that the need for random close associations of the Cy3 and... 

M.J. Rust, M. Bates, X. Zhuang, Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM). Nat. Meth. 3, 793-796 (2006) S.T. Hess, T.P.K. Girirajan, M.D. Mason, Ultra-high resolution imaging by fluorescence photoactivation localization microscopy. Biophys. J. 91(11), 4258-4272 (2006)... 

B. Huang et ah. Three-dimensional super-resolution imaging by stochastic optical reconstruction microscopy. Science 319, 810-813 (2008)... 

Sub-Diffraction-Limit Imaging with Stochastic Optical Reconstruction Microscopy... 

Several far-field light microscopy methods have recently been developed to break the diffraction limit. These methods can be largely divided into two categories (1) techniques that employ spatially patterned illumination to sharpen the point-spread function of the microscope, such as stimulated emission depletion (STED) microscopy and related methods using other reversibly saturable optically linear fluorescent transitions (RESOLFT) [1,2], and saturated structured-illumination microscopy (SSIM) [3], and (2) a technique that is based on the localization of individual fluorescent molecules, termed Stochastic Optical Reconstruction Microscopy (STORM [4], Photo-Activated Localization Microscopy (PALM) [5], or Fluorescence Photo-Activation Localization Microscopy (FPALM) [6]. In this paper, we describe the concept of STORM microscopy and recent advances in the imaging capabilities of STORM. 

Stochastic optical reconstruction microscopy (STORM) and direct STORM (dSTORM), also referred to as ground-state depletion and single-molecule return (GSDIM), use fluorochromes that reversibly cycle between fluorescent (ON) and dark (OFF) states upon exposure to light of specific wavelengths [158, 162-164]. STORM-based ICC (Fig. 3D) relies on the proximity of two fluorochromes attached to an antibody in a specific ratio and at a specific distance (Fig. 3E), whereas dSTORM (Fig. 3E) employs a... 

Direct stochastic optical reconstruction microscopy with standard fluorescent probes. Nat Protoc 6 991-1009... 

Photoartivated localization microscopy (PALM), fluorescence photoartivation localization microscopy (FPALM), and stochastic optical reconstruction microscopy (STORM) are the abbreviations under which this concept is commonly known at present and they all rely on stochastic on/off switching and localization of individual emitters, followed by image reconstruction. 

---