STORM overcomes the diffraction limit by using photo-switchable fluorescent probes to temporally separate the spatially overlapping images of individual molecule. A different subset of fluorescent probes are activated at different times, allowing these fluorophores to be imaged without substantial spatial overlap and to be localized with high precision. Iterating the activation and imaging process allows the position of many fluorescent probes to be determined and a superresolution image to be reconstructed from the positions of the localized probes.

Comparison of conventional diffraction-limited (upper) and STORM (bottom) images of microtubules in a cell. Microtubules are immunolabeled with photoswitchable dyes.

Two-color conventional (Left) and STORM (right) image of microtubules (green) and clathrin-coated pits (red) in a cell. Microtubules and clathrin are immunolabeled with photoswitchable dyes.

Comparison of conventional (Left) and 3D-STORM (right) images of clathrin-coated pits (red) in a cell. Shown on the right is a xy-cross section of a 3D STORM image. Clathrin is immunolabeled with photoswitchable dyes.

Comparison of conventional and 3D STORM image of mitochondria in a cell. The left panel shows the conventional image of the left 1/3 of the cell. The middle panel shows the 3D STORM image of the middle 1/3 of the cell. The z-dimension information is color-coded. The right panel shows the xy cross-section of the 3D STORM image of the right 1/3 of the cell. TOM20 in on mitochondria is immunolabeled with photoswitchable dyes.

3D STORM image of actin in a cell. The z-dimension information is color-coded. Actin is labeled with phalloidin conjugated to photoswitchable dyes.

Comparison of conventional (left) and STORM (right) images of chemical synapses in the brain tissue. The synapses are immunolabeled for presynaptic protein Bassoon (red) and postsynaptic protein Homer1 (green) using photoswitchable dyes.

Comparison of conventional (upper) and STORM (bottom) images of telomere loops. The telomere is labeled by fluorescence in situ hybridization with photoswitchable dyes.

Comparison of conventional (upper) and 3D STORM (lower) images of actin in the axons of neurons. Actin is labeled with phalloidin conjugated to photoswitchable dyes. STORM image revealed a novel periodic, actin-spectrin-based membrane skeleton in axons.

STORM image of a novel periodic, actin-spectrin-based membrane skeleton in axons. This image shows the STORM image of ΒII-spectrin in neuritis immunolabeled with photoswitchable dyes. Except for one neurite in the middle, which is a dendrite, all the other neurities in the image are axons, which exhibit a periodic distribution of ΒII-spectrin.

Two-color STORM images of a novel periodic, actin-spectrin based cytoskeleton in axons. The upper panel shows the STORM image of actin (green) and ΒII-spectrin (magenta). The lower panel shows the STORM image of ΒII-spectrin (green) and adducin (magenta). Actin is labeled with with phalloidin conjugated to photoswitchable dyes. ΒII-spectrin is immunostained labeled with photoswitchable dyes against its C-terminal region, which is situated at the center of the spectrin tetramer. Adducin is immunolabeled with photoswitchable dyes.

STORM image of a novel periodic, actin-spectrin based cytoskeleton in axons. The image shows the STORM image of actin in neurites. The neurite in the middle is an axon, which shows a periodic distribution of actin. Actin is labeled with phalloidin conjugated to photoswitchable dyes.

Comparison of conventional (upper) and STORM (bottom) images of chromatin in the nucleus. A specific locus of the chromatin is labeled by fluorescence in situ hybridization with photoswitchable dyes.

3D-STORM movie of clathrin-coated pits in a cell. Shown are a series of xy-cross sections of a 3D STORM image. Clathrin is immunolabeled with photoswitchable dyes.

3D-STORM movie of ribosomes in a dividing bacterial cell. Shown are a series of xy-cross sections of a 3D STORM image. Ribosome is labeled by fluorescence in situ hybridization with photoswitchable dyes.

3D STORM movie of a chemical synapse in the brain tissue. The synapses are immunolabeled for presynaptic protein Bassoon (red) and postsynaptic protein Homer1 (green) using photoswitchable dyes.

3D STORM movie of a calcium channel, CatSper, in the sperm tail. CatSper is immunolabeled with photoswitchable dyes.

Volumentric STORM movie of dendritic branch immerse in inhibitory synapses. The synapses are immunolabeled for presynaptic proteins (red) and gephrin (green). The neurones express GFP and is immunolabeled using anti-GFP using photoswitchable dyes. The left side of the movie shows an dendritic branch (cyan) amidst synapses, and the right side shows only the synapses assigned to the dendritic branch.

Volumentric STORM movie of a direction-selective retinal ganglion cells with all of its inhibitory input synapses. The synapses are immunolabeled for presynaptic proteins (red) and gephrin (green). The neuron expresses GFP and is immunolabeled using anti-GFP using photoswitchable dyes.