Choosing dyes for cw-STED nanoscopy using self-assembled nanorulers

被引:29
作者
Beater, Susanne [1 ]
Holzmeister, Phil [1 ]
Pibiri, Enrico [1 ]
Lalkens, Birka [1 ]
Tinnefeld, Philip [1 ]
机构
[1] Tech Univ Carolo Wilhelmina Braunschweig, Inst Phys & Theoret Chem, NanoBioSci Grp, D-38106 Braunschweig, Germany
基金
欧洲研究理事会;
关键词
FIELD OPTICAL NANOSCOPY; DNA ORIGAMI; SUPERRESOLUTION MICROSCOPY; SUBDIFFRACTION-RESOLUTION; FLUORESCENCE MICROSCOPY; STIMULATED-EMISSION; PATTERNS; STANDARDS; SHAPES; LIMIT;
D O I
10.1039/c4cp00127c
中图分类号
O64 [物理化学(理论化学)、化学物理学];
学科分类号
070304 ; 081704 ;
摘要
Superresolution microscopy is currently revolutionizing optical imaging. A key factor for getting images of highest quality is - besides a well-performing imaging system - the labeling of the sample. We compared the fluorescent dyes Abberior Star 488, Alexa 488, Chromeo 488 and Oregon Green 488 for use in continuous wave (cw-)STED microscopy in aqueous buffer and in a durable polymer matrix. To optimize comparability, we designed DNA origami standards labeled with the fluorescent dyes including a bead-like DNA origami with dyes focused on one spot and a DNA origami with two marks at a designed distance of similar to 100 nm. Our data show that all dyes are well suited for cw-STED microscopy but that the optimal dye depends on the embedding medium. The precise comparison enabled by DNA origami nanorulers indicates that these structures have matured from the proof-of-concept to easily applicable tools in fluorescence microscopy.
引用
收藏
页码:6990 / 6996
页数:7
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