New near-infrared fluorescent probes with single-photon anti-stokes-shift fluorescence for sensitive determination of pH variances in lysosomes with a double-checked capability

Tzu-Ho Chen, Academia Sinica
Shuwei Zhang, Michigan Technological University
Meghnath Jaishi, Michigan Technological University
Rashmi Adhikari, Michigan Technological University
Jianheng Bi, Michigan Technological University
Mingxi Fang, Michigan Technological University
Shuai Xia, Michigan Technological University
Yibin Zhang, Academia Sinica
Rudy Luck, Michigan Technological University
Ranjit Pati, Michigan Technological University
Hsien-Ming Lee, Academia Sinica
Fen-Tair Luo, Academia Sinica
Ashutosh Tiwari, Michigan Technological University
Haiying Liu, Michigan Technological University

© 2018 American Chemical Society. Publisher's version of record: https://doi.org/10.1021/acsabm.8b00020

Erratum: https://digitalcommons.mtu.edu/michigantech-p/2730/

Abstract

Two near-infrared luminescent probes with Stokes-shift and single-photon anti-Stokes-shift fluorescence properties for sensitive determination of pH variance in lysosomes have been synthesized. A morpholine residue in probe A which serves as a targeting group for lysosomes in viable cells was attached to the fluorophores via a spirolactam moiety while a mannose residue was ligated to probe B resulting in increased biocompatibility and solubility in water. Probes A and B contain closed spirolactam moieties, and show no Stokes-shift or anti-Stokes-shift fluorescence under neutral or alkali conditions. However, the probes incrementally react to pH variance from 7.22 to 2.76 with measurable increases in both Stokes-shift and anti-Stokes-shift fluorescence at 699 nm and 693 nm under 645 nm and 800 nm excitation, respectively. This acid-activated fluorescence is produced by the breaking of the probe spirolactam moiety, which greatly increased overall π-conjugation in the probes. These probes possess upconversion near-infrared fluorescence imaging advantages including minimum cellular photo-damage, tissue penetration, and minimum biological fluorescence background. They display excellent photostability with low dye photobleaching and show good biocompatibility. They are selective and capable of detecting pH variances in lysosomes at excitation at two different wavelengths, i.e., 645 and 800 nm.