RESCue STED 超分辨活细胞成像系统

Abberior RESCue STED超分辨活细胞成像

RESCue = REduction of State transition Cycles

所有的 Abberior STED 超分辨系统, 皆可升级附加 RESCue STED.

全球唯一将超分辨技术实现在活细胞的影像应用

· Pulsed STED超分辨率成像应用在活细胞的长时间成像记录

· 仅需使用一般4%的光计量，解决荧光漂白及对样本光伤害的障碍

RESCueSTED 技术原理

· Abberior独家 RESCue-STED (RESCue = REduction of State transition Cycles )

是 Abberior Pulsed STED 超分辨系统的附加模块, 也是超分辨技术的最新发展, 因可大副降低光剂量, 是唯一可落实在活细胞长时间影像撷取应用, 或者, 荧光容易漂白的样本, 是把超分辨技术应用落实在生物细胞的实用技术., 也是众所期待的最新实用技术. 此RESCue STED 技术已在欧美科研界掀起广泛的使用热潮 !

· 以激光曝晒在样本的光剂量计算, RESCue-STED 仅是  gated-STED 的 4% 而已  !

Abberior RESCue STED的使用效益

· 可随时在任何Abberior Pulsed STED超分辨系统上做升级

· 拥有最高的纯光学分辨率，优于XY 20nm，XYZ（3D） 70X70X70nm

· 可轻易的做活细胞长时间影像撷取

· 可做活细胞荧光动态分析

· 可做活细胞及组织的深层厚度影像撷取

· 可做多色荧光影像撷取

· 荧光不易漂白

RESCue STED技术的基本原理

RESCue (REduction of State transition Cycles) STED is an improved, photon-efficient STED imaging mode that significantly reduces the light dose sent onto your sample without compromising the resolution – STED imaging conditions designed for live-cell STED imaging!

· 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 not make 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

RESCue STED 与其他技术的曝光剂量比较简单的举例, Leica g-STED 的曝光剂量是Abberior RESCue-STED 的25倍 !

使用实务上, 曝光剂量愈少, 对样本(活细胞)的伤害愈小, 荧光也愈不会漂白 !

左边 : RESCue-STED                                              右边 : 传统的 gated-STED

经过 30 次的 RESCue-STED 扫描取像.                   经过 30 次的 gated –CW STED扫描取像,

STED 影像依然清晰可见                                         STED 影像因为重复曝光, 产生严重的荧光漂白, 影像效果极差.

因为几乎没有荧光漂白现象

RESCue STED应用

· RESCue STED 是 Abberior STED 的附加模块, 可以不影响超分辨的光学解析下, 大副降低光辐射剂量, 仅是过去的gated-cw-STED 成像技术的 4% 光剂量而已. 在既有新一代的 Pulsed STED 技术, 有了 RESCue STED 模块, 也仅是使用 20% 的光剂量而已.

· RESCue STED 成像技术, 最适宜于活细胞超分辨成像及 3D-STED 的超分辨立体成像

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. 
Shown are 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

RESCue STED in living yeast cells @ 595 nm.
Shown are Citrine-labelled eisosomes in living yeast cells (sample courtesy of Prof. Dr. S. Jakobs, University Medical Center G?ttingen). Note that only ~4 % of the light dose was applied during image generation (compared to gated cw STED).

相较于 Leica 的 gated CW STED ( TCS SP8 STED ),  Abberior RESCue STED 仅使用小于 4% 的光打到活细胞样本.

所以, 没有光漂白, 光毒害, 可以长时间做活细胞影像撷取

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.

Shown are 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

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