Fluorescent RET-Based Chemosensor Bearing 1,8-Naphthalimide and Styrylpyridine Chromophores for Ratiometric Detection of Hg2+ and Its Bio-Application.

Dyad compound NI-SP bearing 1,8-naphthalimide (NI) and styrylpyridine (SP) photoactive units, in which the N-phenylazadithia-15-crown-5 ether receptor is linked with the energy donor naphthalimide chromophore, has been evaluated as a ratiometric fluorescent chemosensor for mercury (II) ions in living cells. In an aqueous solution, NI-SP selectively responds to the presence of Hg2+ via the enhancement in the emission intensity of NI due to the inhibition of the photoinduced electron transfer from the receptor to the NI fragment. At the same time, the long wavelength fluorescence band of SP, arising as a result of resonance energy transfer from the excited NI unit, appears to be virtually unchanged upon Hg2+ binding. This allows self-calibration of the optical response. The observed spectral behavior is consistent with the formation of the (NI-SP)·Hg2+ complex (dissociation constant 0.13 ± 0.04 µM). Bio-imaging studies showed that the ratio of fluorescence intensity in the 440-510 nm spectral region to that in the 590-650 nm region increases from 1.1 to 2.8 when cells are exposed to an increasing concentration of mercury (II) ions, thus enabling the detection of intracellular Hg2+ ions and their quantitative analysis in the 0.04-1.65 µM concentration range.

Fluorescence turn-on probes for intracellular DNA/RNA distribution based on asymmetric bis(styryl) dyes.

Two bis(styryl) dyes, varying in type of spacer between two mono(styryl) units, were tested for interactions with ct-DNA or cl-RNA. Both compounds showed strong affinity toward ds-DNA/ss-RNA, the binding mode of the interaction is shifting between DNA groove binding to RNA intercalation. Consequently, interaction with DNA shows a stronger flare-up of fluorescence (151 times for dye 1 and 118 times for dye 2) than when binding with RNA (23 times and 36 times correspondingly). The presence of energy transfer in the bis(styryl) system increases the Stokes shift of the dye, so when irradiating the system in the region of 370-380 nm, fluorescence is detected at 610-620 nm. The biological experiments showed that the efficient intracellular fluorescence quench was observed in the DNase digest test suggested that dyes can be applied by recognition of DNA in the presence of RNA molecules.