Alexa Fluor

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The Alexa Fluor family of fluorescent dyes is a series of dyes invented by Molecular Probes, now a part of Thermo Fisher Scientific, and sold under the Invitrogen brand name. Alexa Fluor dyes are frequently used as cell and tissue labels in fluorescence microscopy and cell biology.[1] Alexa Fluor dyes can be conjugated directly to primary antibodies or to secondary antibodies to amplify signal and sensitivity[2] or other biomolecules.

The excitation and emission spectra of the Alexa Fluor series cover the visible spectrum and extend into the infrared.[3] The individual members of the family are numbered according roughly to their excitation maxima (in nm).


History[edit]

Richard and Rosaria Haugland, the founders of Molecular Probes, are well known in biology and chemistry for their research into fluorescent dyes to be used in biological applications. At the time that Molecular Probes was founded, these products were essentially unavailable commercially. A number of now well-known fluorescent dyes came out of the labs at Molecular Probes. Dyes such as Texas Red, Cascade Blue, Oregon Green, Marina Blue and the Alexa Fluor family. Arguably the most famous, the Alexa Fluor family of dyes, were intended to improve upon the properties of previously developed biological fluorescent dye families and solve some of the issues that they possessed. Structurally, the Alexa Fluor dyes are generated through the sulfonation and additional modification of certain well known dye families. In particular, the dye families of coumarin, rhodamine, xanthene (of which the industry-standard fluorescein is a member), and cyanine dyes were used. Sulfonation makes the Alexa Fluor dyes negatively charged and more hydrophilic than their precursors, while additional modification was used to improve the performance in other areas. For example, Alexa Fluor 488, a sulfonated and chemically modified form of fluorescein, was designed to solve the well known issues of rapid photo-bleaching and pH dependent fluorescent intensity common in the industry-standard FITC dye. The Alexa Fluor dyes were named after Alex Haugland, son of Richard and Rosaria Haugland. Molecular Probes was acquired in 2003, by Invitrogen[4] who further worked to expand the Alexa Fluor family adding new dyes to fill gaps in the spectrum. In 2008, Invitrogen became Life Technologies when they merged with Applied Biosystems, bringing the Alexa Fluor family of dyes under the new Life Technologies name. In 2014, Life Technologies was acquired by Thermo Fisher Scientific who revitalized the Invitrogen name and brought the Alexa Fluor family of dyes back under the Invitrogen brand.

Dyes[edit]

  Colour[citation needed] Absorb
(nm)[5]
Emit
(nm)[5]
MM
(g/mol)[citation needed]
ε
(cm−1M−1)[5]
Quantum Yield [6]
Alexa Fluor 350 blue 346 442 410 19,000 -
— 405 violet 401 421 1028 35,000 -
— 430 green 434 541 702 15,000 -
— 488 cyan-green 495 519 643 73,000 0.92
— 500 green 502 525 700 71,000 -
— 514 green 517 542 714 80,000 -
— 532 green 532 554 721 81,000 0.61
— 546 yellow 556 573 1079 112,000 0.79
— 555 yellow-green 555 565 ~1250 155,000 0.1
— 568 orange 578 603 792 88,000 0.69
— 594 orange-red 590 617 820 92,000 0.66
— 610 red 612 628 1172 144,000 -
— 633 Far-red 632 647 ~1200 159,000 -
— 635 Far-red 633 647 - 140,000 -
— 647 Far-red 650 665 1155.06[7] 270,000 0.33
— 660 Near-IR 663 690 ~1100 132,000 0.37
— 680 Near-IR 679 702 ~1150 183,000 0.36
— 700 Near-IR 702 723 ~1400 205,000 0.25
— 750 Near-IR 749 775 ~1300 290,000 0.12
— 790 Near-IR 782 805 - 260,000 -
† = approximate color of the emission spectrum
ε = extinction coefficient

Comparison with other dyes[edit]

While extinction coefficients are known (see the table above), quantum yields and life times are not. Comparisons with other dyes should be considered depending on the conditions (technique) used and performance (signal, background, stability) needed.

The Alexa Fluor series dyes are less pH-sensitive and more photostable than the original dyes (fluorescein, rhodamine, etc.) from which they were synthesized. Brightness comparisons are also generally favorable. Comparisons with other dyes are less consistent, and also even more delicate, depending on the conditions (technique) used. A third party has compared Alexa Fluor 647 dye with Cy5 (similar wavelength), conjugated to DNA.[8]

Similar lines of fluorescent dyes provide an alternative to the Alexa Fluor Dyes (see also the list in Category:Fluorescent dyes).

References[edit]

  1. ^ "Alexa Fluor Dyes Spanning the Visible and Infrared Spectrum". 2007-06-06. Archived from the original on 11 August 2007. Retrieved 2007-08-13. 
  2. ^ "Alexa Fluor Secondary Antibodies". www.thermofisher.com. Retrieved 2017-05-26. 
  3. ^ "The Alexa Fluor Dye Series". Molecular Probes, Inc. 2006-04-06. Archived from the original on 15 August 2007. Retrieved 2007-08-13. 
  4. ^ "Allbusiness.com: Invitrogen Prices $325M Worth Of Notes, Keeps Buying Power.". Retrieved 2010-01-17. 
  5. ^ a b c http://www.lifetechnologies.com/sg/en/home/references/molecular-probes-the-handbook/technical-notes-and-product-highlights/the-alexa-fluor-dye-series.html
  6. ^ "Fluorescence quantum yields (QY) and lifetimes (τ) for Alexa Fluor dyes—Table 1.5". Retrieved 2011-04-26. 
  7. ^ Supporting Information; Esteban, Fink, et al. Fungal recognition is mediated by the association of dectin-1 and galectin-3 in macrophages. Proceedings of the National Academy of Sciences of the United States of America. 2011. Vol 108, no 34, pg 14270-14275. 10.1073/pnas.1111415108
  8. ^ Ballard JL; Peeva VK; deSilva CJ; Lynch JL; Swanson NR (July 2007). "Comparison of Alexa Fluor and CyDye for practical DNA microarray use". Retrieved 23 October 2010. 

External links[edit]