Protons play crucial roles in many physiological and pathological processes, such as receptor-mediated signal transduction, ion transport, endocytosis, homeostasis, cell proliferation, and apoptosis. The urgent demand for pH imaging and measurement in biological systems has incited the development of fluorescent pH probes. Numerous fluorescent probes have been reported, but many lack the abilities needed for biological applications. Hence, the development of new pH probes with better biocompatibility, sensitivity, and site-specificity is still indispensable. This review highlights the recent trends in the development of fluorescent materials as essential tools for tracing pH variations in the biological processes of diverse living systems.

A multifunctional fluorescent sensor based on a cyclen-appended BINOL derivative (R-1) was synthesized and characterized. It can display on-off-type fluorescence change with high selectivity toward Cu(II) among 19 metal ions in 100% aqueous solution. Furthermore, the in situ generated R-1-Cu(II) ensemble could recover the quenched fluorescence upon the addition of sulfide anion resulting in a off-on-type sensing with a detection limit of micromolar range in the same medium. No interference was observed from other biothiols and anions, including GSH, l-Cys, DTT, and sulfates, making it a highly sensitive and selective sulfide probe.

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A reaction-based colorimetric and ratiometric fluorescent probe based on an ICT-strategy for selective detection of H(2)S that exploited the H(2)S-mediated reduction of nitrocompound to amines was explored. And it displayed high selectivity for H(2)S over other relevant reactive sulfur, oxygen, nitrogen species and other anions with more than 120 nm blue shift and the change of emission intensity ratio inducted by H(2)S was over 4750.

A real-time colorimetric and ratiometric fluorescent probe based on modulating the intramolecular charge transfer (ICT) of the coumarin platform for selective detection of sulfite is presented. This reaction based probe utilized the Michael addition to the dicyano-vinyl group with the detection limit of 5.8 × 10(-5) M. The probe displayed a high selectivity for sulfite over other anions and reactive sulfur especially for biothiols including cysteine (Cys), homocysteine (Hcy) and glutathione (GSH), with about 100 nm blue shift and more than 230 times intensity ratios change of the emission spectrum. Meanwhile, it could be easily observed that the probe for sulfite changes from red to pale yellow by the naked eye, and from red to blue under UV lamp immediately after the sulfite is added. To the best of our knowledge, it is the fastest response probe for sulfite ever reported, which could give a colorimetric and ratiometric fluorescent response instantly.

Two rhodamine-based polymers were prepared via free radical polymerization and could serve as lysosome targeting probes with good pH sensitivity. Fluorescence imaging of nude mice displayed a chance for visualization of cancerous tissue in vivo by sensing its acidic microenvironments.

Two mitochondria-targeted real-time probes were presented, which could selectively respond to hypochlorite over other ROS. Meanwhile, the "off-on" probes could be successfully applied in the in vivo imaging of hypochlorite in living mice.

A biotin-rhodamine-TPP conjugate (BiTClO) was reported as a real-time (within 10 s) fluorescent probe for the sensing of mitochondrial hypochlorite with excellent selectivity and high sensitivity (0.21 μM). Moreover, it is the first tumor-specific and mitochondria-targeted fluorescent probe applied in cell imaging.