Nano-imaging with Storm

Abstract

Multicolour, three-dimensional stochastic optical reconstruction microscopy now makes it possible to image cellular structures with near molecular-scale resolution.

Fig. 1. The principle of STORM

For a densely labeled cellular structure shown in a, only a sparse set of fluorescent labels are activated at any given time such that their images (green circles) do not overlap (b). b, The position of each activated fluorophore (red crosses) is then determined from its image. c, After a sufficient number of fluorophores have been localized, a high resolution image is constructed by plotting the measured positions. Modified with permission from ref. . © 2009 Annual Reviews.

Fig. 2. STORM imaging of cells with photo-switchable dyes and fluorescent proteins

a, STORM image of microtubules in a BS-C-1 cell. The microtubules are immunostained with antibodies that are labeled with photo-switchable Alexa Fluor 647. b and c, Conventional (b) and STORM (c) images corresponds to the boxed region in a. d and e, Conventional (d) and STORM (e) images of vimentin in a BS-C-1 cell. The vimentin filaments are labeled with a photo-switchable fluorescent protein mEos2.

Fig. 3. Multicolour STORM

The microtubules and clathrin-coated pits in a BS-C-1 cell are immunostained with Cy2-Alexa 647-labeled and Cy3-Alexa 647-labeled antibodies, respectively. a, Conventional fluorescence image. b, STORM image of the same area. c, Magnified view of the boxed region in b. Parts b and c adapted with permission from ref. . © 2007 AAAS.

Figure 4. Three-dimensional STORM

a, Conventional image of clathrin-coated pits in a BS-C-1 cell. Clathrin is immuno-stained with Cy3-Alexa 647-labeled antibodies. b, x-y cross-section of the corresponding 3D STORM image. c, Serial x-y and x-z cross-sections of a coated pit. d, An x-y and x-z cross section presented in 3D perspective. Parts a–d from ref. . © 2008 AAAS.