Tags

Type your tag names separated by a space and hit enter

Masked rhodamine dyes of five principal colors revealed by photolysis of a 2-diazo-1-indanone caging group: synthesis, photophysics, and light microscopy applications.
Chemistry 2014; 20(41):13162-73C

Abstract

Caged rhodamine dyes (Rhodamines NN) of five basic colors were synthesized and used as "hidden" markers in subdiffractional and conventional light microscopy. These masked fluorophores with a 2-diazo-1-indanone group can be irreversibly photoactivated, either by irradiation with UV- or violet light (one-photon process), or by exposure to intense red light (λ∼750 nm; two-photon mode). All dyes possess a very small 2-diazoketone caging group incorporated into the 2-diazo-1-indanone residue with a quaternary carbon atom (C-3) and a spiro-9H-xanthene fragment. Initially they are non-colored (pale yellow), non-fluorescent, and absorb at λ=330-350 nm (molar extinction coefficient (ε)≈10(4) M(-1) cm(-1)) with a band edge that extends to about λ=440 nm. The absorption and emission bands of the uncaged derivatives are tunable over a wide range (λ=511-633 and 525-653 nm, respectively). The unmasked dyes are highly colored and fluorescent (ε=3-8×10(4) M(-1) cm(-1) and fluorescence quantum yields (ϕ)=40-85% in the unbound state and in methanol). By stepwise and orthogonal protection of carboxylic and sulfonic acid groups a highly water-soluble caged red-emitting dye with two sulfonic acid residues was prepared. Rhodamines NN were decorated with amino-reactive N-hydroxysuccinimidyl ester groups, applied in aqueous buffers, easily conjugated with proteins, and readily photoactivated (uncaged) with λ=375-420 nm light or intense red light (λ=775 nm). Protein conjugates with optimal degrees of labeling (3-6) were prepared and uncaged with λ=405 nm light in aqueous buffer solutions (ϕ=20-38%). The photochemical cleavage of the masking group generates only molecular nitrogen. Some 10-40% of the non-fluorescent (dark) byproducts are also formed. However, they have low absorbance and do not quench the fluorescence of the uncaged dyes. Photoactivation of the individual molecules of Rhodamines NN (e.g., due to reversible or irreversible transition to a "dark" non-emitting state or photobleaching) provides multicolor images with subdiffractional optical resolution. The applicability of these novel caged fluorophores in super-resolution optical microscopy is exemplified.

Authors+Show Affiliations

NanoBiophotonics Department, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen (Germany), Fax: (+49) 551-201-2505. vbelov@gwdg.de.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25196166

Citation

Belov, Vladimir N., et al. "Masked Rhodamine Dyes of Five Principal Colors Revealed By Photolysis of a 2-diazo-1-indanone Caging Group: Synthesis, Photophysics, and Light Microscopy Applications." Chemistry (Weinheim an Der Bergstrasse, Germany), vol. 20, no. 41, 2014, pp. 13162-73.
Belov VN, Mitronova GY, Bossi ML, et al. Masked rhodamine dyes of five principal colors revealed by photolysis of a 2-diazo-1-indanone caging group: synthesis, photophysics, and light microscopy applications. Chemistry. 2014;20(41):13162-73.
Belov, V. N., Mitronova, G. Y., Bossi, M. L., Boyarskiy, V. P., Hebisch, E., Geisler, C., ... Hell, S. W. (2014). Masked rhodamine dyes of five principal colors revealed by photolysis of a 2-diazo-1-indanone caging group: synthesis, photophysics, and light microscopy applications. Chemistry (Weinheim an Der Bergstrasse, Germany), 20(41), pp. 13162-73. doi:10.1002/chem.201403316.
Belov VN, et al. Masked Rhodamine Dyes of Five Principal Colors Revealed By Photolysis of a 2-diazo-1-indanone Caging Group: Synthesis, Photophysics, and Light Microscopy Applications. Chemistry. 2014 Oct 6;20(41):13162-73. PubMed PMID: 25196166.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Masked rhodamine dyes of five principal colors revealed by photolysis of a 2-diazo-1-indanone caging group: synthesis, photophysics, and light microscopy applications. AU - Belov,Vladimir N, AU - Mitronova,Gyuzel Yu, AU - Bossi,Mariano L, AU - Boyarskiy,Vadim P, AU - Hebisch,Elke, AU - Geisler,Claudia, AU - Kolmakov,Kirill, AU - Wurm,Christian A, AU - Willig,Katrin I, AU - Hell,Stefan W, Y1 - 2014/09/05/ PY - 2014/04/29/received PY - 2014/9/9/entrez PY - 2014/9/10/pubmed PY - 2015/6/3/medline KW - bioconjugation KW - diazo compounds KW - fluorescence KW - photolysis KW - rhodamines SP - 13162 EP - 73 JF - Chemistry (Weinheim an der Bergstrasse, Germany) JO - Chemistry VL - 20 IS - 41 N2 - Caged rhodamine dyes (Rhodamines NN) of five basic colors were synthesized and used as "hidden" markers in subdiffractional and conventional light microscopy. These masked fluorophores with a 2-diazo-1-indanone group can be irreversibly photoactivated, either by irradiation with UV- or violet light (one-photon process), or by exposure to intense red light (λ∼750 nm; two-photon mode). All dyes possess a very small 2-diazoketone caging group incorporated into the 2-diazo-1-indanone residue with a quaternary carbon atom (C-3) and a spiro-9H-xanthene fragment. Initially they are non-colored (pale yellow), non-fluorescent, and absorb at λ=330-350 nm (molar extinction coefficient (ε)≈10(4) M(-1) cm(-1)) with a band edge that extends to about λ=440 nm. The absorption and emission bands of the uncaged derivatives are tunable over a wide range (λ=511-633 and 525-653 nm, respectively). The unmasked dyes are highly colored and fluorescent (ε=3-8×10(4) M(-1) cm(-1) and fluorescence quantum yields (ϕ)=40-85% in the unbound state and in methanol). By stepwise and orthogonal protection of carboxylic and sulfonic acid groups a highly water-soluble caged red-emitting dye with two sulfonic acid residues was prepared. Rhodamines NN were decorated with amino-reactive N-hydroxysuccinimidyl ester groups, applied in aqueous buffers, easily conjugated with proteins, and readily photoactivated (uncaged) with λ=375-420 nm light or intense red light (λ=775 nm). Protein conjugates with optimal degrees of labeling (3-6) were prepared and uncaged with λ=405 nm light in aqueous buffer solutions (ϕ=20-38%). The photochemical cleavage of the masking group generates only molecular nitrogen. Some 10-40% of the non-fluorescent (dark) byproducts are also formed. However, they have low absorbance and do not quench the fluorescence of the uncaged dyes. Photoactivation of the individual molecules of Rhodamines NN (e.g., due to reversible or irreversible transition to a "dark" non-emitting state or photobleaching) provides multicolor images with subdiffractional optical resolution. The applicability of these novel caged fluorophores in super-resolution optical microscopy is exemplified. SN - 1521-3765 UR - https://www.unboundmedicine.com/medline/citation/25196166/Masked_rhodamine_dyes_of_five_principal_colors_revealed_by_photolysis_of_a_2_diazo_1_indanone_caging_group:_synthesis_photophysics_and_light_microscopy_applications_ L2 - https://doi.org/10.1002/chem.201403316 DB - PRIME DP - Unbound Medicine ER -