Abberior® Dyes for super-resolution microscopy applications

BioFiles Vol. 9

Microscopic methods in life sciences are of tremendous importance for visualization cellular and tissue structures. The development of new microscopy concepts has overcome the resolution barrier given by the diffraction limit, enabling a resolution limit down to about 10 nm. The visualization of cellular structures and molecular interactions can reveal new understanding in biological processes. Due to tremendous efforts in the development of super-resolved fluorescence microscopy, The Nobel Prize in Chemistry for 2014 was awarded to Eric Betzig, Stefan W. Hell, and to William E. Moerner.

These super-resolution microscopy principles are based on several technological approaches. Conventional light microscopy enables a resolution limit of about 250 nm in the x- and y- direction and 450 – 700 nm in the z –direction. Super-resolution techniques have overcome the resolution-limit (point-spread function) at least by a factor of 2. The resolution of super-resolution microscopy depends on the number of points that can be resolved on the structure of interest. Crucial for a successful super-resolution imaging is the choice of fluorescent probe. Brightness and high-contrast ratio between the states are important. In most super-resolution methods, the states of the probe must be controllable, reversible or irreversible, and switchable between a light or a dark state. Depending on the super-resolution method, further photo-physical criteria the probe must be fulfilled. Established techniques include the following:

  • STED (Stimulated emission depletion)
  • GSDIM (Ground State Depletion)
  • PALM (Photoactivated localization microscopy)
  • STORM (Stochastic optical reconstruction microscopy)
  • RESOLFT (reversible saturable optical (flurorescence) transitions)

 

3 color STED image of primary hippocampal neurons imaged with the Abberior® Instruments Expert Line STED microscope

Three-color STED image of primary hippocampal neurons imaged with the Abberior® Instruments Expert Line STED microscope. Note the characteristic ~190 nm beta II spectrin periodicity along distal axons (green), which is only visible in the STED image. Labeled structures and dyes: beta II Spectrin (green, Abberior® STAR580), Bassoon (red, Abberior® STAR635P), Actin cytoskeleton (blue, Phalloidin, Oregon Green 488). Sample was prepared by Elisa D’Este at MPIBPC, Göttingen.

We offer the superior series of Abberior® dyes that are especially designed and tested for super-resolution microscopy such as STED, RESOLFT, PALM, STORM, GSDIM and others. Abberior® STAR, Abberior® CAGE, Abberior® FLIP, Abberior® RSFP – the specific requirements of the super-resolution techniques are served with dedicated dye series. These Dyes are developed and produced by Abberior® GmbH. Stefan Hell is it’s Co-founder.

Benefits

  • Optimized for brightness and low background
  • Optimized switching behavior being the key for super-resolution
  • All markers are tested for different super-resolution methods
    • Abberior® STAR for STED, confocal and epifluorescence imaging
    • Abberior® CAGE & FLIP for PALM, STORM and GSDIM
  • Abberior® dyes are recommended by microscope vendors
  • Proprietary, IP protected products
  • Detailed characteristics of the dyes provided, e.g. optimal STED wavelength

Super-resolution microscopy is dependent on fluorescent labels. Manufactured by Abberior®, the STAR, CAGE and FLIP dyes as well as RSFPs are exceptionally bright and photostable and provide optimized photoswitching for RESOLFT and PALM/STORM imaging. They are the only commercially available dyes that are tailored specifically to the needs of super-resolution microscopy.

Abberior® dyes are also well suited for confocal microscopy, epifluorescence imaging and single molecule applications. Fluorescence applications that depend on a good signal-to-noise ratios and low background cam benefit from Abberior® dyes.

Two subunits of the nuclear pore complex were immunolabelled using antibodies against gp210 and antibodies with multiple specificities (PAN4/5) and secondary antibodies coupled to Abberior® STAR580 and Abberior ® STAR635P

Confocal and STED images. Two subunits of the nuclear pore complex were immunolabeled using antibodies against gp210 and antibodies with multiple specificities (PAN4/5) and secondary antibodies coupled to Abberior® STAR580 and Abberior® STAR635P. Note that gp210 is localized in an eight-fold symmetric structure at the rim of the nuclear pore complex. Imaged with the Abberior® Instruments STEDYCON (compact line).


Product Table: Overview of Abberior® Dyes
 

Dyes Description Absorption Maximum, λabs Extinction Coefficient, ε Fluorescence Maximum, λfl Recommended STED Cat No. NHS Cat No. Maleimide
Abberior® CAGE 500 for single-molecule switching microscopy (e.g. PALM, STORM, GSDIM) 230, 299, 340 nm (non-activated, PBS, pH 7.4)
511 nm (photoactivated, PBS, pH 7.4)
50,000 M-1cm-1
(photoactivated, PBS, pH 7.4)
525 nm (PBS, pH 7.4) 595-615 nm 44254 92546
Abberior® CAGE 532 for single-molecule switching microscopy (e.g. PALM, STORM, GSDIM) 237, 302, 350 nm (non-activated, PBS, pH 7.4)
533 nm (photoactivated, PBS, pH 7.4)
82,000 M-1cm-1
(photoactivated, PBS, pH 7.4)
541 nm (PBS, pH 7.4) 610-640 nm   95705
Abberior® CAGE 552 for single-molecule switching microscopy (e.g. PALM, STORM, GSDIM) 231, 308, 350 nm (non-activated, PBS, pH 7.4)
552 nm (photoactivated, PBS, pH 7.4)
66,000 M-1cm-1
(photoactivated, PBS, pH 7.4)
574 nm (PBS, pH 7.4) 650-670 nm 94822 92545
Abberior® CAGE 590 for single-molecule switching microscopy (e.g. PALM, STORM, GSDIM) 262, 325, 351 nm (non-activated, PBS, pH 7.4)
595 nm (photoactivated, PBS, pH 7.4)
75,000 M-1cm-1
(photoactivated, PBS, pH 7.4)
615 nm (PBS, pH 7.4) 685-715 nm 77958 no
Abberior® FLIP 565 for single-molecule switching microscopy (e.g. PALM, STORM, GSDIM) 314 nm (closed form, PBS, pH 7.4)
566 nm (open form, PBS, pH 7.4)
47,000 M-1cm-1
(open form, PBS pH, 7.4)
580 nm (open form, PBS, pH 7.4) - 79189 92544
Abberior® STAR 440SXP for long Stokes STED and 2-color STED application 432 nm (PBS, pH 7.4) 33,000 M-1cm-1
(PBS, pH 7.4)
511 nm (PBS, pH 7.4) 590-620 nm 68221 38361
Abberior® STAR 470SXP for long Stokes STED and 2-color STED application 467 nm (PBS, pH 7.4) 29,000 M-1cm-1
(PBS, pH 7.4)
598 nm (PBS, pH 7.4) 740-770 nm 94716 no
Abberior® STAR 488 for STED application 503 nm (PBS, pH 7.4) 65,000 M-1cm-1
(PBS, pH 7.4)
524 nm (PBS, pH 7.4) 585-605 nm 61048 no
Abberior® STAR 512 for STED application 511 nm (PBS, pH 7.4) 85,000 M-1cm-1
(PBS, pH 7.4)
530 nm (PBS, pH 7.4) 590-620 nm 38922 03004
Abberior® STAR 580 for STED application 587 nm (PBS, pH 7.4) 85,000 M-1cm-1
(PBS, pH 7.4)
607 nm (PBS, pH 7.4) 690-720 nm 38377 no
Abberior® STAR 635 for STED application 635 nm (PBS, pH 7.4) 110,000 M-1cm-1
(PBS, pH 7.4)
655 nm (PBS, pH 7.4) 740-770 nm 30558 96013
Abberior® STAR 635P for STED application 638 nm (PBS, pH 7.4) 120,000 M-1cm-1
(PBS, pH 7.4)
651 nm (PBS, pH 7.4) 740-770 nm 07679 no

 

For North America, we also provide a series of antibody labeled Abberior® conjugates.
 

Cat No. Description Availability Pack size
53647 Anti-Rabbit IgG-Abberior® CAGE 635 antibody produced in goat US only 500 µg
30483 Anti-Mouse IgG-Abberior® CAGE 635 antibody produced in goat US only 500 µg
54287 Anti-Rabbit IgG-Abberior® CAGE 590 antibody produced in goat US only 500 µg
53364 Anti-Mouse IgG-Abberior® CAGE 590 antibody produced in goat US only 500 µg
40544 Anti-Rabbit IgG-Abberior® CAGE 552 antibody produced in goat US only 500 µg
53165 Anti-Mouse IgG-Abberior® CAGE 552 antibody produced in goat US only 500 µg
53601 Anti-Rabbit IgG-Abberior® CAGE 532 antibody produced in goat US only 500 µg
52996 Anti-Mouse IgG-Abberior® CAGE 532 antibody produced in goat US only 500 µg
41155 Anti-Rabbit IgG-Abberior® CAGE 500 antibody produced in goat US only 500 µg
52953 Anti-Mouse IgG-Abberior® CAGE 500 antibody produced in goat US only 500 µg
52283 Anti-Mouse IgG-Abberior® STAR  RED antibody produced in goat US only 500 µg
53399 Anti-Rabbit IgG-Abberior® STAR 635P antibody produced in goat US only 500 µg
41699 Anti-Rabbit IgG-Abberior® STAR RED antibody produced in goat US only 500 µg
41348 Anti-Rabbit IgG-Abberior® STAR 635 antibody produced in goat US only
(available soon)
500 µg
40734 Anti-Mouse IgG-Abberior® STAR 635 antibody produced in goat US only 500 µg
53654 Anti-Rabbit IgG-Abberior® STAR 600 antibody produced in goat US only 500 µg
41367 Anti-Rabbit IgG-Abberior® STAR 580 antibody produced in goat US only 500 µg
52597 Anti-Mouse IgG-Abberior® STAR 600 antibody produced in goat US only 500 µg
52403 Anti-Mouse IgG-Abberior® STAR 580 antibody produced in goat US only 500 µg
52932 Anti-Rabbit IgG-Abberior® STAR 520SXP antibody produced in goat US only 500 µg
41372 Anti-Mouse IgG-Abberior® STAR  520SXP antibody produced in goat US only 500 µg
52269 Anti-Mouse IgG-Abberior® STAR 512 antibody produced in goat US only 500 µg
00289 Anti-Rabbit IgG-Abberior® STAR 512 antibody produced in goat US only 500 µg
52944 Anti-Rabbit IgG-Abberior® STAR 488 antibody produced in goat US only 500 µg
53366 Anti-Mouse IgG-Abberior® STAR 488 antibody produced in goat US only 500 µg
52187 Anti-Mouse IgG-Abberior® STAR 470SXP antibody produced in goat US only 500 µg
41324 Anti-Rabbit IgG-Abberior® STAR 470SXP antibody produced in goat US only 500 µg
41860 Anti-Rabbit IgG-Abberior® STAR 440SXP antibody produced in goat US only 500 µg
41738 Anti-Mouse IgG-Abberior® STAR 440SXP antibody produced in goat US only 500 µg

 

 

General References

  1. Leica Microsystems recommendations for 2-color applications.
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  10. S. Li, et al. "High Speed Optical Nanoscopy by Stimulated Emission Depletion (STED) with Galvo Mirrors", roc. SPIE 8911, International Symposium on Photoelectronic Detection and Imaging 2013: Micro/Nano Optical Imaging Technologies and Applications (2013)
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  17. S. Rocha, H. et al. Photoswitchable Fluorescent Proteins for Superresolution Fluorescence Microscopy Circumventing the Diffraction Limit of Light in Fluorescence Spectroscopy and Microscopy (Eds.: Y. Engelborghs, A. J. W. G. Visser), Humana Press, Vol. pp. 793-812 (2013)
  18. J. V. Chacko, et al. Probing Cytoskeletal Structures by Coupling Optical Superresolution and AFM Techniques for a Correlative Approach. Cytoskeleton, 70(11), pp. 729–740, DOI: 10.1002/cm.21139 (2013)
  19. I.-H. Wang, et al. Tracking Viral Genomes in Host Cells and Single Molecule Resolution. Cell Host Microbe, 14(4), pp. 468–480, DOI: 10.1016/j.chom.2013.09.004 (2013)
  20. H. Schill, et al. 4-Trifluoromethyl-Substituted Coumarins with Large Stokes Shifts: Synthesis, Bioconjugates, and Their Use in Super-Resolution Fluorescence Microscopy. Chem. Eur. J., 19(49), pp. 16556–16565, DOI: 10.1002/chem.201302037 (2013)
  21. Y. Wang, et al. Time-Gated Stimulated Emission Depletion Nanoscopy. Opt. Eng., 52(9), 093107, DOI: 10.1117/1.OE.52.9.093107 (2013)
  22. M. V. Sednev, et al. Carborhodol: A New Hybrid Fluorophore Obtained by Combination of Fluorescein and Carbopyronine Dye Cores. Bioconjugate Chem., 24(4), pp 690–700, DOI: 10.1021/bc3006732 (2013)
  23. The Nobel Prize in Chemistry 2014". Nobelprize.org. Nobel Media AB 2014. Web. 14 Oct 2014.