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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.


Related in: MedlinePlus

Luminescence intensity decay curves (A) and the average luminescence lifetime changes (B) of TRP-NO reacted 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.
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fig4: Luminescence intensity decay curves (A) and the average luminescence lifetime changes (B) of TRP-NO reacted 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.

Mentions: Because the intramolecular LRET also dramatically affects the luminescence lifetime of energy donor and acceptor luminophores in the probe, the luminescence decay curves of TRP-NO upon reaction with different concentrations of NO were investigated. As shown in Fig. 4A, with the increase in NO concentration, the luminescence decay rate of TRP-NO at 540 nm increased gradually, and the average luminescence lifetime (decoded from the luminescence decay curve) of TRP-NO decreased gradually from 484.3 μs to 48.7 μs upon reaction with different concentrations of NO (Fig. 4B), which provides a ∼10-fold contrast window for the detection of NO when luminescence lifetime is used as a signal. The results imply that the probe TRP-NO could be also used for the luminescence lifetime imaging of NO in vitro and in vivo. The multiple-signaling feature of TRP-NO enables the NO detection to be carried out both with ratiometric time-gated mode and lifetime mode, which affords much convenience for the accurate quantitative detection of NO in complicated biological samples.


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.
Luminescence intensity decay curves (A) and the average luminescence lifetime changes (B) of TRP-NO reacted 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.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig4: Luminescence intensity decay curves (A) and the average luminescence lifetime changes (B) of TRP-NO reacted 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.
Mentions: Because the intramolecular LRET also dramatically affects the luminescence lifetime of energy donor and acceptor luminophores in the probe, the luminescence decay curves of TRP-NO upon reaction with different concentrations of NO were investigated. As shown in Fig. 4A, with the increase in NO concentration, the luminescence decay rate of TRP-NO at 540 nm increased gradually, and the average luminescence lifetime (decoded from the luminescence decay curve) of TRP-NO decreased gradually from 484.3 μs to 48.7 μs upon reaction with different concentrations of NO (Fig. 4B), which provides a ∼10-fold contrast window for the detection of NO when luminescence lifetime is used as a signal. The results imply that the probe TRP-NO could be also used for the luminescence lifetime imaging of NO in vitro and in vivo. The multiple-signaling feature of TRP-NO enables the NO detection to be carried out both with ratiometric time-gated mode and lifetime mode, which affords much convenience for the accurate quantitative detection of NO in complicated biological samples.

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.


Related in: MedlinePlus