A sensitive and rapid "off-on" fluorescent probe for the detection of esterase and its application in evaluating cell status and discrimination of living cells and dead cells.

The discrimination of living and dead cells shows great importance in the development of biology, pathology, medicine, and pharmacology research. Herein, we synthesized a simple benzothiazole-based probe, EP, which was characterized via1H NMR (hydrogen nuclear magnetic resonance) spectroscopy, 13C NMR (carbon nuclear magnetic resonance) spectroscopy and HRMS (high-resolution mass spectroscopy). The fluorescence changes in response to esterase were characterized via fluorescence spectroscopy. EP exhibited a 70-fold fluorescence enhancement in the presence of esterase and possessed a very low limit of detection (4.73 × 10-5 U mL-1). EP also showed high selectivity to esterase compared to other biological species. Bright fluorescence appeared in living cells, which was activated by esterase when incubated with EP. In paraformaldehyde or H2O2 pretreated cells, the fluorescence became very weak since esterase became inactive in these cells. In summary, the EP probe can monitor esterase activity both in vitro and in living cells and can be used to evaluate the health status of cells and discriminate living and dead cells effectively.

A GSH Fluorescent Probe with a Large Stokes Shift and Its Application in Living Cells.

Intracellular GSH is the most abundant non-protein biothiol and acts as a central antioxidant to defend against aging toxins and radicals. Meanwhile abnormal level of intracellular GSH concentration is directly related to some diseases. In this case, detecting intracellular GSH rapidly and sensitively is of great significance. We synthesize a simple fluorescent probe (named GP) which can discriminate GSH from Cys (cysteine) or Hcy (homocysteine) and presents a 50-fold fluorescence increasing. The response time of GP to GSH was only 5 min and the product GO (the product of GP after reacting with GSH) after reacting with GSH possesses a larger Stokes shift for 135 nm than that in reported work. Probe GP can detect intracellular effectively and shows obvious yellow fluorescence. Briefly, probe GP can detect intracellular GSH rapidly and effectively both in vitro and in living cells.

Fluorescence turn-on detection of Sn2+ in live eukaryotic and prokaryotic cells.

Sn(2+) is usually added to toothpaste to prevent dental plaque and oral disease. However, studies of its physiological role and bacteriostatic mechanism are restricted by the lack of versatile Sn(2+) detection methods applicable to live cells, including Streptococcus mutans. Here we report two Sn(2+) fluorescent probes containing a rhodamine B derivative as a fluorophore, linked via the amide moiety to N,N-bis(2-hydroxyethyl)ethylenediamine (R1) and tert-butyl carbazate group (R2), respectively. These probes can selectively chelate Sn(2+) and show marked fluorescence enhancement due to the ring open reaction of rhodamine induced by Sn(2+) chelation. The probes have high sensitivity and selectivity for Sn(2+) in the presence of various relevant metal ions. Particularly, both R1 and R2 can target lysosomes, and R2 can probe Sn concentrations in lysosomes with rather acidic microenvironment. Furthermore, these two probes have low toxicity and can be used as imaging probes for monitoring Sn(2+) not only in live KB cells (eukaryotic) but also in Streptococcus mutans cells (prokaryotic), which is a useful tool to study the physiological function of Sn(2+) in biological systems.