New from Abberior Instruments: RESCue STED!

Fig7 tubu red

Abberior Instruments RESCue STED is a STED imaging mode dedicated to live-cell imaging with saving up to 96% of the light dose without compromising the resolution.  

RESCue STED is an improved, photon-efficient STED imaging mode that significantly reduces the light dose send onto your sample without compromising the resolution – STED imaging conditions designed for live-cell STED imaging!

RESCue stands for REduction of State transition Cycles:

  • RESCue avoids unnecessary excitation and de-excitation cycles and thus reduces photobleaching of any fluorescent marker
  • The reduction in excitation and de-excitation cycles is particularly beneficial for volume imaging with 3D STED as well as for time-lapse STED imaging
  • RESCue STED applies excitation and STED light only at positions, where fluorescent markers (i.e. the target structure) is present. It shuts-off the lasers at positions where exposure would make no sense (i.e. where no markers are found)
  • The resolution of the image is maintained although the light dose is reduced down to 4% compared to gated cw STED imaging and down to 20% compared to pulsed STED imaging
  • RESCue STED is a highly photon efficient way of superresolution imaging while conventional STED applies the same light dose to any pixel no matter if markers (i.e. a target) are present or not
  • RESCue can be applied with similar benefits also to confocal imaging

Same resolution but only 4% of the typical light dose used – Do you think this is impossible? Please check out our images and movies:

The basic idea of RESCue is shown in the following graphic:

 Fig1 RESCue

Fig2

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Fig3 RESCue STED ON

The blue color indicates, at which sample positions the excitation and the STED laser were shut-off in the RESCue STED imaging mode at the lower threshold. Note that the blue positions correlate with the positions where no marker (i.e. target structure) is present. Sample: Vero cells labelled with antibodies against nuclear pore complex subunits in the central channel of the complex and secondary antibodies coupled to Abberior STAR RED.  

Fig4 redRESCue

RESCue STED @ 775 nm facilitates to record multiple STED images at the same position.
Sample: Vero cells labelled with antibodies against nuclear pore complex subunits in the central channel of the complex and secondary antibodies coupled to Abberior STAR RED.  

Fig5 greenRESCue

 

RESCue STED @ 595 nm facilitates to record multiple STED images at the same position.
Sample: Vero cells labelled with antibodies against nuclear pore complex subunits in the central channel of the complex and secondary antibodies coupled to Alexa Fluor 488.  

 figure 7

RESCue STED @ 775 nm.
Sample: Vero cells labelled with antibodies against ß-tubulin and secondary antibodies coupled to Abberior STAR 635P.  

 figure 8

RESCue STED @ 595 nm.
Sample: Vero cells labelled with phalloidin coupled to Oregon Green 488.

Fig6 Yeast

 

RESCue STED in living yeast cells @ 595 nm.
Sample: Citrine labeled eisosomes in living yeast cells. (sample courtesy of Prof. Dr. S. Jakobs, University Medical Center Göttingen). Note that only ~4% of the light dose was applied during image generation (compared to gated CW STED).  

figure 9

RESCue STED @ 595 nm in living mammalian (Vero) cells expressing MAP2-Citrine (sample courtesy of Prof. Dr. S. Jakobs, University Medical Center Göttingen).  

NUP 3D

RESCue 3D STED @ 775 nm
A 3D image stack of complete mammalian nucleus was recorded with RESCue 3D STED and conventional STED microscopy. The upper part of the figure shows XZ sections of a 3D STED image stack. The lower part shows three planes (XY sections) of the 3D STED image stack. The position of the individual XY planes is labeled by A,B,C in the top images. Sample: Vero cells labelled with antibodies against nuclear pore complex subunits in the central channel of the complex and secondary antibodies coupled to Abberior STAR RED.  

Fig11 3Dstack

RESCue 3D STED @ 775 nm.
A 3D image stack of complete mammalian nucleus was recorded with RESCue 3D STED and conventional STED microscopy. Shown are XZ sections. Sample: Vero cells labelled with antibodies against lamin and secondary antibodies coupled to Abberior STAR 635P. 
Note that RESCue 3D STED enabled us to acquire the complete stack.  

 

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