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Development of a novel lysosome-targetable time-gated luminescence probe for ratiometric and luminescence lifetime detection of nitric oxide in vivo † † Electronic supplementary information (ESI) available: Experimental details for the syntheses of TRP-Tb 3+ and TRP-NO , and supplementary figures. See DOI: 10.1039/c6sc03667h Click here for additional data file.

View Article: PubMed Central - PubMed

ABSTRACT

Trp-notrp-notrp-notrp-notrp-notrp-notrp-no: Rapid, multiplexed, sensitive and specific identification and quantitative detection of nitric oxide (NO) are in great demand in biomedical science. Herein, a novel multifunctional probe based on the intramolecular LRET (luminescence resonance energy transfer) strategy, , was designed for the highly sensitive and selective ratiometric and luminescence lifetime detection of lysosomal NO. Before reaction with NO, the emission of the rhodamine moiety in is switched off, which prevents the LRET process, so that the probe emits only the long-lived Tb3+ luminescence. However, upon reaction with NO, accompanied by the turn-on of rhodamine emission, the LRET from the Tb3+-complex moiety to rhodamine moiety occurs, which results in a remarkable increase of the rhodamine emission and decrease of the Tb3+ emission. After the reaction, the intensity ratio of the rhodamine emission to the Tb3+ emission, I565/I540, was found to be 28.8-fold increased, and the dose-dependent enhancement of the I565/I540 value showed a good linearity upon the increase of NO concentration. In addition, a dose-dependent luminescence lifetime decrease was distinctly observed between the average luminescence lifetime of the probe and NO concentration, which provides a ∼10-fold contrast window for the detection of NO. These unique properties allowed to be conveniently used as a time-gated luminescence probe for the quantitative detection of NO using both luminescence intensity ratio and luminescence lifetime as signals. The applicability of for ratiometric time-gated luminescence imaging of NO in living cells was investigated. Meanwhile, dye co-localization studies confirmed a quite precise distribution of in lysosomes by confocal microscopy imaging. Furthermore, the practical applicability of was demonstrated by the visualization of NO in Daphnia magna. All of the results demonstrated that could serve as a useful tool for exploiting and elucidating the function of NO at sub-cellular levels with high specificity, accuracy and contrast.

No MeSH data available.


UV-vis absorption spectrum changes of TRP-NO (15 μM) after reaction with different concentrations of NO (0, 5, 10, 20, 40, 80, 120, 160, 200, 240, 320, 400 μM) in 0.05 M PBS buffer of pH 7.4 for 50 min (the inserted photographs show colors of the solutions under visible light).
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fig3: UV-vis absorption spectrum changes of TRP-NO (15 μM) after reaction with different concentrations of NO (0, 5, 10, 20, 40, 80, 120, 160, 200, 240, 320, 400 μM) in 0.05 M PBS buffer of pH 7.4 for 50 min (the inserted photographs show colors of the solutions under visible light).

Mentions: The UV-vis spectrum changes of TRP-NO in the presence of different concentrations of NO were further investigated in 0.05 M PBS buffer at pH 7.4. As shown in Fig. 3, after reaction with NO, the absorption of TRP-NO at 557 nm was increased significantly. At the same time, the solution's color was remarkably changed from colorless to pink, implying that the spirolactam ring of the CTMR moiety in the probe was opened after the reaction.


Development of a novel lysosome-targetable time-gated luminescence probe for ratiometric and luminescence lifetime detection of nitric oxide in vivo † † Electronic supplementary information (ESI) available: Experimental details for the syntheses of TRP-Tb 3+ and TRP-NO , and supplementary figures. See DOI: 10.1039/c6sc03667h Click here for additional data file.
UV-vis absorption spectrum changes of TRP-NO (15 μM) after reaction with different concentrations of NO (0, 5, 10, 20, 40, 80, 120, 160, 200, 240, 320, 400 μM) in 0.05 M PBS buffer of pH 7.4 for 50 min (the inserted photographs show colors of the solutions under visible light).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5384565&req=5

fig3: UV-vis absorption spectrum changes of TRP-NO (15 μM) after reaction with different concentrations of NO (0, 5, 10, 20, 40, 80, 120, 160, 200, 240, 320, 400 μM) in 0.05 M PBS buffer of pH 7.4 for 50 min (the inserted photographs show colors of the solutions under visible light).
Mentions: The UV-vis spectrum changes of TRP-NO in the presence of different concentrations of NO were further investigated in 0.05 M PBS buffer at pH 7.4. As shown in Fig. 3, after reaction with NO, the absorption of TRP-NO at 557 nm was increased significantly. At the same time, the solution's color was remarkably changed from colorless to pink, implying that the spirolactam ring of the CTMR moiety in the probe was opened after the reaction.

View Article: PubMed Central - PubMed

ABSTRACT

Trp-notrp-notrp-notrp-notrp-notrp-notrp-no: Rapid, multiplexed, sensitive and specific identification and quantitative detection of nitric oxide (NO) are in great demand in biomedical science. Herein, a novel multifunctional probe based on the intramolecular LRET (luminescence resonance energy transfer) strategy, , was designed for the highly sensitive and selective ratiometric and luminescence lifetime detection of lysosomal NO. Before reaction with NO, the emission of the rhodamine moiety in is switched off, which prevents the LRET process, so that the probe emits only the long-lived Tb3+ luminescence. However, upon reaction with NO, accompanied by the turn-on of rhodamine emission, the LRET from the Tb3+-complex moiety to rhodamine moiety occurs, which results in a remarkable increase of the rhodamine emission and decrease of the Tb3+ emission. After the reaction, the intensity ratio of the rhodamine emission to the Tb3+ emission, I565/I540, was found to be 28.8-fold increased, and the dose-dependent enhancement of the I565/I540 value showed a good linearity upon the increase of NO concentration. In addition, a dose-dependent luminescence lifetime decrease was distinctly observed between the average luminescence lifetime of the probe and NO concentration, which provides a ∼10-fold contrast window for the detection of NO. These unique properties allowed to be conveniently used as a time-gated luminescence probe for the quantitative detection of NO using both luminescence intensity ratio and luminescence lifetime as signals. The applicability of for ratiometric time-gated luminescence imaging of NO in living cells was investigated. Meanwhile, dye co-localization studies confirmed a quite precise distribution of in lysosomes by confocal microscopy imaging. Furthermore, the practical applicability of was demonstrated by the visualization of NO in Daphnia magna. All of the results demonstrated that could serve as a useful tool for exploiting and elucidating the function of NO at sub-cellular levels with high specificity, accuracy and contrast.

No MeSH data available.