A near-infrared fluorescent probe based on a FRET rhodamine donor linked to a cyanine acceptor for sensitive detection of intracellular pH alternations

Yibin Zhang, Michigan Technological University
Jianheng Bi, Michigan Technological University
Shuai Xia, Michigan Technological University
Wafa Mazi, Michigan Technological University
Shulin Wan, Michigan Technological University
Logan Mikesell, Michigan Technological University
Rudy Luck, Michigan Technological University
Haiying Liu, Michigan Technological University

© 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). Publisher’s version of record: https://doi.org/10.3390/molecules23102679


A fluorescence resonance energy transfer (FRET)-based near-infrared fluorescent probe (B+) for double-checked sensitive detection of intracellular pH changes has been synthesized by binding a near-infrared rhodamine donor to a near-infrared cyanine acceptor through robust C-N bonds via a nucleophilic substitution reaction. To demonstrate the double-checked advantages of probe B+, a near-infrared probe (A) was also prepared by modification of a near-infrared rhodamine dye with ethylenediamine to produce a closed spirolactam residue. Under basic conditions, probe B+ shows only weak fluorescence from the cyanine acceptor while probe A displays nonfluorescence due to retention of the closed spirolactam form of the rhodamine moiety. Upon decrease in solution pH level, probe B+ exhibits a gradual fluorescence increase from rhodamine and cyanine constituents at 623 nm and 743 nm respectively, whereas probe A displays fluorescence increase at 623 nm on the rhodamine moiety as acidic conditions leads to the rupture of the probe spirolactam rings. Probes A and B+ have successfully been used to monitor intracellular pH alternations and possess pKa values of 5.15 and 7.80, respectively.