Abstract
Fluorescent/phosphorescent dual-emissive polymers or hybrids consisting of both fluorophore and phosphor have been used as self-calibrating probes and imaging reagents for cellular molecular oxygen. Oxygen selectively quenches the phosphorescence and the fluorescence serves as an internal reference. The phosphorescence/fluorescence ratio is used as a quantitative indicator of oxygen content. In wavelength-ratiometric probes, the fluorophore and phosphor are designed to emit at different wavelengths. It is easy to achieve spectral separation, but the phosphorescence/fluorescence ratio fluctuates due to the difference in the absorption and scattering of light at different wavelengths by biological samples. Herein we reported a lifetime-ratiometric luminescent polymeric probe where the fluorophore and phosphor emitted at the same wavelength. Spectral separation was achieved based on the difference in their excited-state lifetimes via time-resolved luminescence analysis and imaging. The probe exhibited a phosphorescence lifetime of about 931 ns with a phosphorescence/fluorescence ratio of 4.49 in deaerated aqueous buffer. The lifetime was shortened to 251 ns and the ratio decreased to 1.08 in oxygen saturated solution because of phosphorescence quenching. The utilization of the probe for quantitative oxygen sensing and mapping in living HeLa cells was demonstrated using calibration curves obtained from fixed cells.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.