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Matrix metalloproteinase-2 (MMP-2) is a very important biomarker in blood. Presently, sensitive and selective determination of MMP-2 directly in blood samples is still a challenging job because of the high complexity of the sample matrix. In this work, we reported a new homogeneous biosensor for MMP-2 based on fluorescence resonance energy transfer (FRET) from upconversion phosphors (UCPs) to carbon nanoparticles (CNPs). A polypeptide chain (NH(2)-GHHYYGPLGVRGC-COOH) comprising both the specific MMP-2 substrate domain (PLGVR) and a π-rich motif (HHYY) was designed and linked to the surface of UCPs at the C terminus. The FRET process was initiated by the π-π interaction between the peptide and CNPs, which thus quenched the fluorescence of the donor. Upon the cleavage of the substrate by the protease at the amide bond between Gly and Val, the donor was separated from the acceptor while the π-rich motif stayed on the acceptor. As a result, the fluorescence of the donor was restored. The fluorescence recovery was found to be proportional to the concentration of MMP-2 within the range from 10-500 pg/mL in an aqueous solution. The quantification limit of this sensor was at least 1 order of magnitude lower than that of other reported assays for MMP-2. The sensor was used to determine the MMP-2 level directly in human plasma and whole blood samples with satisfactory results obtained. Owing to the hypersensitivity of the method, clinical samples of only less than 1 μL were needed for accurate quantification, which can be meaningful in MMP-2-related clinical and bioanalytical applications.

A ratiometric two-photon probe was designed to visualize HClO levels in tissues, revealing the generation of HClO in the wound-repairing process of mice for the first time.

The dynamics of the room-temperature ionic liquid (RTIL) 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide (BmimNTf) were investigated with two-dimensional infrared (2D IR) vibrational echo spectroscopy and polarization selective pump-probe (PSPP) experiments. The CN stretch frequency of a modified Bmim cation (2-SeCN-Bmim), in which a SeCN moiety was substituted onto the C-2 position of the imidazolium ring, was used as a vibrational probe. A major result of the 2D IR experiments is the observation of a long time scale structural spectral diffusion component of 600 ps in addition to short and intermediate time scales similar to those measured for selenocyanate anion (SeCN) dissolved in BmimNTf. In contrast to 2-SeCN-Bmim, SeCN samples its inhomogeneous line width nearly an order of magnitude faster than the complete structural randomization time of neat BmimNTf liquid (870 ± 20 ps) measured with optical heterodyne-detected optical Kerr effect (OHD-OKE) experiments. The orientational correlation function obtained from PSPP experiments on 2-SeCN-Bmim exhibits two periods of restricted angular diffusion (wobbling-in-a-cone) followed by complete orientational randomization on a time scale of 900 ± 20 ps, significantly slower than observed for SeCN but identical within experimental error to the complete structural randomization time of BmimNTf. The experiments indicate that 2-SeCN-Bmim is sensitive to local motions of the ionic region that influence the spectral diffusion and reorientation of small, anionic, and neutral molecules as well as significantly slower, longer-range fluctuations that are responsible for complete randomization of the liquid structure.

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Hydrogen persulfide and polysulfide (H2S(n)) are newly discovered intracellular reactive species considered to have high protein S-sulfhydration efficiency. The detection of H2S(n) in living systems is essential for studying their functions but is quite challenging. In this work, we report a two-photon excited fluorescent probe, QS(n), capable of tracking H2S(n) in living organisms. QS(n) exhibited turn-on two-photon fluorescence response upon reaction with H2S(n). With a favorable photophysical property, high specificity, and low cytotoxicity, QS(n) was able to recognize exogenous H2S(n) in living cells. More importantly, it realized for the first time the visualization of endogenous H2S(n) generated in cells overexpressing cystathionine β-synthase and cystathionine γ-lyase, the enzymes responsible for producing endogenous H2S(n). Taking advantage of two-photon microscopy, the probe was also applied to achieve H2S(n) detection in zebrafish embryos and to observe H2S(n) distribution in living organisms.

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