Super-Resolution Microscopy Dyes
CF® Dyes for multicolor super-resolution microscopy
Recent publications comparing synthetic dyes for super-resolution imaging have shown that CF® dyes give the best performance for multiple methods. The superior brightness, photostability, and photochemical switching properties of certain CF® dyes are ideal for 3-D SIM, 3-D STORM, and other super-resolution techniques.
Biotium's CF®405M has been found to be the brightest and most photostable short wavelength fluorescent dye for SIM. Several CF® dyes spanning the visible red, far-red, and near-infrared spectra have been validated for STORM, including three color imaging with CF®568, CF®647, and CF®680. See Lehmann et al. 2015, and a full list of references on the next page. Biotium offers a wide selection of CF® dye labeled antibodies, including single-label secondaries with low degree of labeling, which is reported to be optimal for STORM1. We also offer other conjugates, reactive dyes, and labeling kits.
Figure 1. CF®568 (left) produces better images than Cy®3b (right) in 3-D STORM microscopy.
Fixed cells were stained with mouse anti-tubulin antibody followed by dye-conjugated antimouse secondary antibodies. See Figure 2 legend for imaging conditions. Dye molecules were photoswitched and imaged using a 560 nm laser; a 405 nm laser was used to assist dye reactivation to the emitting state
Figure 2. Comparison of conventional wide-field microscopy (left) with STORM (right) using CF® dye conjugates.
Fixed cells were stained with mouse anti-tubulin antibody followed by CF® dye conjugated anti-mouse secondary antibody (top row: CF®647, middle row: CF®660C, bottom row: CF®680). For STORM, samples were sealed in buffer that contained 5%(w/v) glucose, 100 mM cysteamine, 0.8 mg/mL glucose oxidase, and 40 μg/mL catalase, in Tris-HCl (pH 7.5). Samples were imaged on a Nikon Ti-Eclipse w/ PFS microscope with a CFI Plan Apo Lambda 100x oil objective. Dye molecules were photoswitched and imaged using a 647 nm laser; a 405 nm laser was used to assist dye reactivation to the emitting state. Emission was collected with an Andor iXon Ultra 897 EMCCD camera for a total of 100,000 frames per image at a frame rate of 110 Hz.
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