Inspired by our unique RESCue concept DyMIN STED further reduces sample areas where maximum STED power is applied. This reduces the overall bleaching and enables to increase the reachable optical resolution with STED microscopy.
A second conceptual ingredient of DyMIN STED is MINFIELD STED, a co-development between Stefan Hell and coworkers and Abberior Instruments.
DyMIN STED complements an existing list of intelligent STED illumination schemes:
DyMIN STED opens up optical resolutions significantly below 30 nm on the full field of view. DyMIN STED imaging of DNA origami structures at high signal-to-noise ratio clearly separates two fluorescent spots of 30 nm distance (GATTAquant DNA origami sample).
DyMIN STED very clearly resolves spectrin rings with superior signal-to-noise ratio. Primary hippocampal neurons (22 days in vitro) show the characteristic ~192 nm betaII spectrin periodicity along distal axons. Dye: Abberior STAR 635P, Excitation: 635 nm, STED: 775 nm. Sample with courtesy from Elisa D’Este (MPIbpc).
Easy3D DyMIN STED significantly reduces bleaching and thus enables the acquisition of a volume stack with an isotropic resolution of ~85 nm. Primary hippocampal neurons (22 days in vitro), betaII spectrin. Dye: Abberior STAR 635P, Excitation: 635 nm, STED: 775 nm. Sample with courtesy from Elisa D’Este (MPIbpc). Scale bars 200 nm.
Easy3D DyMIN STED clearly resolves two nuclear pore complex subunits (NPC, green) on the inner and outer side of the nuclear membrane (red). Targets: nup153 (green), lamin (red). Dyes: Alexa 594, Abberior STAR 635P, Excitation: 561nm, 635 nm, STED: 775 nm.
After acquisition with DyMIN STED no bleaching is observed. Vimentin labeled with Oregon Green 488; Excitation: 488 nm, STED: 595 nm.
After acquisition with DyMIN STED significantly lower bleaching is observed. Nuclear pore complex protein (nup153) labeled with Oregon Green 488; Excitation: 488 nm, STED: 595 nm.
A 3D image stack of the mammalian nucleus was recorded with easy3D DyMIN STED and conventional STED microscopy. Shown are xz sections of the stack and a confocal image after the stack-acqusition, highlighting the remarkable bleaching reduction of DyMIN STED. Note that DyMIN STED enabled us to acquire the complete stack with superior resolution and signal. Shown are Vero cells labelled with antibodies against nuclear pore complex protein (nup153).
Video: DyMIN STED in living fibroplast cell. Note that 117 consecutive STED images have been recorded without fluorescence signal loss.