Accessing lanthanide-based, in situ illuminated optical turn-on probes by modulation of the antenna triplet state energy.

Luminescent lanthanides possess ideal properties for biological imaging, including long luminescent lifetimes and emission within the optical window. Here, we report a novel approach to responsive luminescent Tb(iii) probes that involves direct modulation of the antenna excited triplet state energy. If the triplet energy lies too close to the 5D4 Tb(iii) excited state (20 500 cm-1), energy transfer to 5D4 competes with back energy transfer processes and limits lanthanide-based emission. To validate this approach, a series of pyridyl-functionalized, macrocyclic lanthanide complexes were designed, and the corresponding lowest energy triplet states were calculated using density functional theory (DFT). Subsequently, three novel constructs L3 (nitro-pyridyl), L4 (amino-pyridyl) and L5 (fluoro-pyridyl) were synthesized. Photophysical characterization of the corresponding Gd(iii) complexes revealed antenna triplet energies between 25 800 and 30 400 cm-1 and a 500-fold increase in quantum yield upon conversion of Tb(L3) to Tb(L4) using the biologically relevant analyte H2S. The corresponding turn-on reaction can be monitored using conventional, small-animal optical imaging equipment in presence of a Cherenkov radiation emitting isotope as an in situ excitation source, demonstrating that antenna triplet state energy modulation represents a viable approach to biocompatible, Tb-based optical turn-on probes.

Investigating the Time Response of an Optical pH Sensor Based on a Polysiloxane-Polyethylene Glycol Composite Material Impregnated with a pH-Responsive Triangulenium Dye.

Determining the time it takes a sensor to report a change in the concentration of its target analyte may appear to be an easy task, but it is not. The dynamic characteristic of a sensor is determined by all components in the sensor system and the hydrodynamics of the sample. Here, the dynamic properties of an optical pH sensor were determined using the IUPAC-recommended activity step method in experimental setups that can determine sensor-limited response times longer than 5 s. In order to do so, experimental setups for the injection and for the dipping method of determining the sensor time response were developed, tested, and shown to be able to determine time-response curves with 1 s time resolution. This time resolution is shown to be sufficient for determining dynamic characterization of this optical pH sensor. The sensor chemistry-limited time-response curves were analyzed using curve fitting. It was found that the optode response time is limited by diffusion of protons within the sensor material when the proton concentration is reduced and limited by diffusion from the bulk to the boundary layer at the optode surface when proton concentration is increased. The latter is dependent on the magnitude of the change in analyte concentration and cannot be reported as a single response time. The investigation of the time response of the optical pH sensor reveals detailed information of the sensor chemistry, but does not yield a single response time of the sensor capable of describing the dynamic sensor characteristics of the optical pH sensor system.

Tuning the p Ka of a pH Responsive Fluorophore and the Consequences for Calibration of Optical Sensors Based on a Single Fluorophore but Multiple Receptors.

Since Sørensen and Bjerrum defined the pH scale, we have relied on two methods for determining pH, the colorimetric or the electrochemical. For pH electrodes, calibration is easy as a linear response is observed in the interesting pH range from 1 to ∼12. For colorimetric sensors, the response follows the sigmoidal Bjerrum diagram of an acid-base equilibrium. Thus, calibration of colorimetric sensors is more complex. Here, seven pH responsive fluorescent dyes based on the same diazaoxatriangulenium (DAOTA) fluorophore linked to varying receptor groups were prepared. Photoinduced electron transfer (PeT) quenching from appended aniline or phenol receptors generated the pH response of the DAOTA dyes, and the position of the p Ka value of the dye was tuned using the Hammett relationship as a guideline. The fluorescence intensity of the dyes in a sol-gel matrix environment was measured as a function of pH in universal buffer, and it was found that the dyes behave as perfect pH responsive probes under these conditions. The response of optical pH sensors is nonlinear and was found to be limited to 2-3 pH units for a precision of 0.01 pH unit. As sensors with a broader sensitivity range can be achieved by mixing multiple dyes with different p Ka values, mixtures of dyes in solution were investigated, and a broad range pH sensor with a precision of 0.006 pH units over a range of 3.6 pH units was demonstrated. Further, approximating the sensor response as linear was considered, and a limiting precision for this approach was determined. As the responses of the pH responsive DAOTA dyes were found to be ideally sigmoidal and as the six dyes were shown to have p Ka values scattered over a range from ∼2 to ∼9, this allows for design of a broad range optical pH sensor in the pH range from 1 to 10. This hypothesis was tested using quaternary mixtures of the different DAOTA dyes, and these were found to behave as a direct sum of the individual components. Thus, while linear calibration is limited to a precision of 0.02 in a range of 2-3 pH units, calibration using ideal sigmoidal functions is possible in the range of 1-10 with a precision better than 0.01, and as good as 0.002 pH units.

Optical Chemical Sensor Using Intensity Ratiometric Fluorescence Signals for Fast and Reliable pH Determination.

Optical pH sensors enable noninvasive monitoring of pH, yet in pure sensing terms, the potentiometric method of measuring pH is still vastly superior. Here, we report a full spectrometer-based optical pH sensor system consisting of sensor chemistry, hardware, and software that for the first time is capable of challenging the performance of an electrode-based pH meter in specific applications such as biopharmaceutical process monitoring and in single-use bioproduction. A highly photostable triangulenium fluorophore emitting at 590 nm was immobilized in an organically modified silicon matrix that allows for fast time-response by rapid diffusion of water in and out of the resulting composite polymer deposited on a polycarbonate substrate. Fluctuations from the fiber optical sensor hardware have been reduced by including a highly photostable terrylene-based reference dye emitting at 660 nm, thus enabling intensity-based ratiometric readouts. The dyes were excited by 505 nm light from a light emitting diode. The sensor was operational within a pH range of 4.6-7.6, and was characterized and demonstrated to have properties that are comparable to those of commercial pH electrodes considering time-response ( t90 < 90 s), precision (0.03 pH-units), and drift.

The critical shoulder angle is associated with osteoarthritis in the shoulder but not rotator cuff tears: a retrospective case-control study.

BACKGROUND: In 2013 Moor et al introduced the concept of the critical shoulder angle (CSA) and suggested that an abnormal CSA was a leading factor in development of rotator cuff tear (RCT) and osteoarthritis (OA) of the shoulder. This study assessed whether the CSA was associated with RCT and OA and tested the inter- and intrarater reliability of the CSA when measuring RCT and OA. MATERIALS AND METHODS: The study was performed as a retrospective case-control study. The cases comprised 2 groups: 97 patients with RCT and 87 patients with OA. The controls were matched 3:1, by age and sex, from a population of 795 patients with humeral fractures. The CSA was measured as described by Moor et al. Analysis of the relation with CSA for RCT and OA was done by logistic regression. Models were fitted separately for RCT and OA and used the controls matched to the respective cases. Inter- and intrarater reliability was determined by measuring the intraclass correlation coefficient and minimal detectable change. RESULTS: The mean CSA was 33.9° in the RCT group and 33.6° in the matched control group. The odds ratio for developing RCT for people with a CSA above 35° was 1.12 (P = .63). The mean CSA in the OA group was 31.1° and in the matched control group 33.3°. The odds ratio for developing OA for people with a CSA below 30° was 2.25 (P = .002). The CSA measurements showed strong intra- and inter-rater reliability, with intraclass correlation coefficient values above 0.92 and minimal detectable change values below 0.4°. CONCLUSIONS: This study did not find any association between CSA and RCT but did show association between CSA and OA, with a 2.25 odds ratio of developing OA given the patient had a CSA below 30°. The results do not support the suggested praxis of shaving away the lateral border of the acromion to make the CSA smaller because it might increase the risk of developing OA without decreasing the risk of developing RCT. The CSA measurements showed excellent intra- and inter-rater reliability.

Generating multiple-pulse bursts for enhanced fluorescence detection.

The signal-to-background ratio is the limiting factor for fluorescence based detection, sensing, and imaging. A typical background signal will include direct scattering of excitation and Raman scattering of the sample as well as autofluorescence from the sample and additives. To improve the signal-to-background ratio, fluorophores of high brightness and/or high concentration of the fluorophores need to be used. Most of the background is instantaneous and short-lived (picosecond to nanosecond time scale), and using long-lived fluorescence probes combined with time-gated detection allows for significant suppression of unwanted background. Unfortunately, this approach requires substantial sacrifice of the probe signal in order to sufficiently filter the background unless the fluorescence lifetime of the probe is very long. However, long lived probes like ruthenium bipyridyl have relatively low brightness compared to probes that have shorter, 10-30 ns fluorescence lifetimes.We recently presented an approach based on bursts of multiple pulses that allowed for high probe signal amplification using long-lived ruthenium based probe (Ru) and an 80 MHz repetition-rate laser excitation. Unfortunately, Ru represents an extreme case for probe lifetime, and a probe with a shorter lifetime of 20 ns will require excitation from a pulsed source with much higher repetition rate to significantly enhance its signal. Such high repetition rates are not possible to generate with most of today's available electronics. In this report we present new approaches to optimize and generate bursts of pulses with high repetition rate within the burst and no need for new or improved electronics. The high repetition rates originate from a low-repetition source and are highly tunable. We demonstrate that a burst of 2-10 pulses spaced 3 ns apart (corresponding to a 'burst repetition rate' of 330 MHz) allows for high signal enhancement of the 20 ns probe over the sub-nanosecond/nanosecond background. Such an approach can be applied for any sensing format, allowing much higher sensitivity for detection. Since the energy of a single pulse is spread over a few pulses in the burst, the fluorophore's photostability also improves.

Long-lived bright red emitting azaoxa-triangulenium fluorophores.

The fluorescence lifetimes of most red emitting organic probes are under 4 nanoseconds, which is a limiting factor in studying interactions and conformational dynamics of macromolecules. In addition, the nanosecond background autofluorescence is a significant interference during fluorescence measurements in cellular environment. Therefore, red fluorophores with longer lifetimes will be immensely helpful. Azaoxa-triangulenium fluorophores ADOTA and DAOTA are red emitting small organic molecules with high quantum yield, long fluorescence lifetime and high limiting anisotropy. In aqueous environment, ADOTA and DAOTA absorption and emission maxima are respectively 540 nm and 556 nm, and 556 nm and 589 nm. Their emission extends beyond 700 nm. Both probes have the limiting anisotropy between 0.36-0.38 at their absorption peak. In both protic and aprotic solvents, their lifetimes are around 20 ns, making them among the longest-lived red emitting organic fluorophores. Upon labeling of avidin, streptavidin and immunoglobulin their absorption and fluorescence are red-shifted. Unlike in free form, the protein-conjugated probes have heterogeneous fluorescence decays, with the presence of both significantly quenched and unquenched populations. Despite the presence of significant local motions due to a flexible trimethylene linker, we successfully measured both intermediate nanosecond intra-protein motions and slower rotational correlation times approaching 100 ns. Their long lifetimes are unaffected by the cell membrane (hexadecyl-ADOTA) and the intra-cellular (DAOTA-Arginine) localization. Their long lifetimes also enabled successful time-gating of the cellular autofluorescence resulting in background-free fluorescence lifetime based images. ADOTA and DAOTA retain a long fluorescence lifetime when free, as protein conjugate, in membranes and inside the cell. Our successful measurements of intermediate nanosecond internal motions and long correlations times of large proteins suggest that these probes will be highly useful to study slower intra-molecular motions and interactions among macromolecules. The fluorescence lifetime facilitated gating of cellular nanosecond autofluorescence should be of considerable help in in vitro and in vivo applications.

Reversible recruitment and emission of DO3A-derived lanthanide complexes at ligating molecular films on gold.

The recruitment of DO3A-derived lanthanide complexes by ligation to isophthalic acid and catechol-modified gold surfaces, and their resulting sensitization, is reported herein. Predictably pH-dependent surface recruitment is associated with the expected fingerprint europium and terbium emission characteristics. The intensity of the lanthanide luminescence scales exponentially with spacer length, indicating a strong quenching interaction between the lanthanide and the gold surface. The switchable catechol oxidation state provides a means of electrochemically triggering the release of prior ligated complexes.

Obtaining enhanced circular dichroism in [4]heterohelicenium analogues.

Electronic circular dichroism in the three helicenium analogues dimethoxyquinacridinium (DMQA(+)), dimethoxychromenoacridinium (DMCA(+)), and dimethoxychromenoxanthenium (DMCX(+)) were investigated in vacuum with time-dependent density functional theory methods using the CAM-B3LYP functional with the basis set 6-311++G**. The systems were thoroughly studied by designing derivatives with several different electron-donating and -withdrawing substituents while at the same time keeping the net charge of the molecule either positive, neutral, or negative. Fifty-six derivatives were examined, and we identify a superior substitution pattern that is found to be independent of the bridging atoms and gives a rotational strength close to 90 × 10(-40) esu(2) cm(2) for DMQA, DMCA, and DMCX. The optimal system shows promising applications as a chromophore because it has a highly allowed primary electronic transition with an angle between the electronic and magnetic transition dipole moments close to 50° and its chiroptical response is thereby limited only by the magnetic transition.

Racemization mechanisms and electronic circular dichroism of [4]heterohelicenium dyes: a theoretical study.

Triarylmethylium cations with the three rings linked by two bridging groups constitute a special class of [4]heterohelicenium dyes that combine high configurational stability with the optical properties of classic dye compounds. The racemization barriers and electronic circular dichroism of seven [4]heterohelicenium analogues are investigated with density functional theory to consider different design strategies. The racemization barriers are examined with the B3LYP functional utilizing the basis set 6-31+G* with respect to bridging heteroatoms in the helicenium motif and with different bulky substituents in the helix pitch. The racemization barriers of the [4]heterohelicenium are found to be highly dependent on the relative size of the bridging atom. Nucleophilic attack at the carbenium ion leads to formation of center adducts for which the racemization barriers are found to be lowered by 8-12 kJ/mol. This finding of a nucleophilic racemization catalysis may be rationalized by the loss of conjugation upon formation of an sp(3)-hybridized carbon in the center adducts. The effect of the heteroatom substitution and center adduct formation is reflected in the electronic properties of the compound calculated with the Coulomb-attenuated method CAM-B3LYP with the basis set 6-311++G**. The excitation energies are found to be highly dependent on the twisting of the helicenium framework and only weakly influenced by the electronic nature of the bridging substituent. The electronic circular dichroism is evaluated, as the rotational strength is found to be highly dependent on both the overall molecular structure and substitution pattern but with no simple correlation between structure and circular dichroism (CD) response. The calculations reveal that the magnitude of rotational strength in most cases is dominated by the angle between the electronic and magnetic transition dipole moments. Finally, it is found that computational screening of many different structures and substituents might be needed to find target structures with maximized rotational strength.

Spectroscopic properties of curcumin: orientation of transition moments.

Curcumin, a naturally occurring yellow-orange pigment with potent antioxidant and antitumor properties, has been attracting researchers from a wide range of fields including chemistry, spectroscopy, biology, and medicine. Ultrafast excited-state processes such as solvation and excited-state intramolecular hydrogen atom transfer (ESIHT) make curcumin an attractive agent for photodynamic therapy. In this report we present studies of linear dichroism and fluorescence anisotropy in oriented and isotropic media. The results show transition moments (long wavelength absorption and emission) oriented along the long molecular axis. Comparison of linear dichroism and excitation anisotropy in oriented and isotropic media suggests that excited-state intramolecular hydrogen atom transfer is probably associated with intramolecular conformational changes that can be constrained in highly stretched poly(vinyl alcohol) (PVA) film.

Synthesis and optical properties of trioxatriangulenium dyes with one and two peripheral amino substituents.

Several derivatives of two new dye systems, with one or two dialkylamino donor groups attached to resonant positions at the periphery of a trioxatriangulenium ion, were synthesized. The mono- and bis-dialkylamino trioxatriangulenium salts (A(1)-TOTA(+) and A(2)-TOTA(+)) were prepared from methoxy-substituted triphenylmethylium (TPM) compounds by aromatic nucleophilic substitution with secondary amines and subsequent intramolecular ring closure. The optical properties of the new triangulenium dyes and their TPM precursors were investigated and compared to those of known TPM and xanthenium dyes. The optical properties were found to be dependent on symmetry and charge localization in the conjugated framework. The trioxatriangulenium dye with two amino groups (A(2)-TOTA(+)) was found to be a strong fluorophore with properties as a blue-shifted rhodamine B. The mono-substituted compound (A(1)-TOTA(+)) was found to be only weakly fluorescent. We assign the weak fluorescence of A(1)-TOTA(+) to an efficient but reversible formation of a nonfluorescent conformation in the excited state, favored by the large degree of charge localization in this dye with only one donor group.

Synthesis of super stable triangulenium dye.

The super stabilized tris(dialkylamino)-azadioxatriangulenium carbenium ion (A-ADOTA(+)) 4 has been synthesized and characterized. As a result of extended delocalization of the positive charge in the planar triangulenium system and the presence of several strongly electron-donating groups, the A-ATOTA(+) ion displays extreme cation stability with a pK(R+) value close to 25. Furthermore the A-ATOTA(+) ion shows excellent dye properties with intense absorption at 458 nm and high fluorescence quantum yield.

Structure of the buried metal-molecule interface in organic thin film devices.

By use of specular X-ray reflectivity (XR) the structure of a metal-covered organic thin film device is measured with angstrom resolution. The model system is a Langmuir-Blodgett (LB) film, sandwiched between a silicon substrate and a top electrode consisting of 25 A titanium and 100 A aluminum. By comparison of XR data for the five-layer Pb2+ arachidate LB film before and after vapor deposition of the Ti/Al top electrode, a detailed account of the structural damage to the organic film at the buried metal-molecule interface is obtained. We find that the organized structure of the two topmost LB layers (approximately 5 nm) is completely destroyed due to the metal deposition.

Complexation between methyl viologen (paraquat) bis(hexafluorophosphate) and dibenzo[24]crown-8 revisited.

Paraquat bis(hexafluorophosphate) undergoes stepwise dissociation in acetone. All three species-the neutral molecule, and the mono- and dications-are represented significantly under the experimental conditions typically used in host-guest binding studies. Paraquat forms at least four host-guest complexes with dibenzo[24]crown-8. They are characterized by both 1:1 and 1:2 stoichiometries, and an overall charge of either zero (neutral molecule) or one (monocation). The monocationic 1:1 host-guest complex is the most abundant species under typical (0.5-20 mM) experimental conditions. The presence of the dicationic 1:1 host-guest complex cannot be excluded on the basis of our experimental data, but neither is it unambiguously confirmed to be present. The two confirmed forms of paraquat that do undergo complexation-the neutral molecule and the monocation-exhibit approximately identical binding affinities toward dibenzo[24]crown-8. Thus, the relative abundance of neutral, singly, and doubly charged pseudorotaxanes is identical to the relative abundance of neutral, singly, and doubly charged paraquat unbound with respect to the crown ether in acetone. In the specific case of paraquat/dibenzo[24]crown-8, ion-pairing does not contribute to host-guest complex formation, as has been suggested previously in the literature.

Enhanced Fluorescence Emission of Me-ADOTA by Self-Assembled Silver Nanoparticles on a Gold Film.

We report a multi-fold enhancement of the fluorescence of methyl-azadioxatriangulenium chloride (Me-ADOTA*Cl) in PVA deposited on a 50 nm thick gold mirror carrying an evaporation induced self-assembly of colloidal silver nanoparticles (Ag-SACs). The average measured increase in fluorescence emission of about 50-fold is accompanied by hot spots with a local enhancement in brigthness close to 200. The long lifetime of the dye allows for the first direct determination of the correlation between the enhancement of emission intensity and the decrease in fluorescence lifetime. The Ag-SACs surface preparation and observed enhancements are highly reproducible. We believe that these robust plasmonic surfaces will find use in sensing platforms for ultrasensitive detection.

Synthesis of novel amphiphilic azobenzenes and X-ray scattering studies of their Langmuir monolayers.

We report a simple synthetic route to novel symmetrical alkylated and acylated amphiphilic 4,4'-diaminoazobenzene dyes, with their optical axis perpendicular to the amphiphilic direction of the molecule. Three different substitution patterns are reported, two of which are highly amphiphilic. At the air-water interface, the amphiphilic azobenzenes form noncrystalline but stable Langmuir films that display an unusual reversible monolayer collapse close to 35 mN/m. The structures and phase transitions were studied by X-ray reflectivity (XR) and grazing-incidence X-ray diffraction, both utilizing synchrotron radiation. Compression beyond the collapse point does not change the XR data, showing that the film is unchanged at the molecular level, even at areas less than half of that of the collapse. This leads to the conclusion that few macroscopic collapse sites are responsible for reversibly removing large amounts of material from the interface.