Ultrafast Molecular Rotor Research Articles

Reply to Comment on "Emissive H-Aggregates of an Ultrafast Molecular Rotor: A Promising Platform for Sensing Heparin".

Reply to Comment on "Emissive H-Aggregates of an Ultrafast Molecular Rotor: A Promising Platform for Sensing Heparin".

Comment on "Emissive H-Aggregates of an Ultrafast Molecular Rotor: A Promising Platform for Sensing Heparin".

For heparin sensing, Mudliar and Singh developed fluorescence and absorption spectroscopic approaches by utilizing emissive H-aggregates of thioflavin T (ThT) formed upon heparin binding. It has been proposed that the methods work not only in pure aqueous solution but also in complex biological media such as human serum. However, the optical features used to detect and quantify heparin are very sensitive to the ionic strength of the solution and completely vanish at 1.45 mM NaCl. Curiously, the authors were able to determine the heparin content of 1% serum samples containing the same level of electrolyte. In addition, the experimental conditions employed for heparin detection in serum samples were substantially modified, reducing the optical path length from 1 to 0.1 cm and increasing the dye concentration by an unknown measure. ThT shows a concentration-dependent tendency for aqueous aggregation, which markedly modifies its absorption and fluorescence properties. The authors have failed to verify that spectral characteristics of the ThT-heparin system observed in pure aqueous medium remain unchanged at higher dye concentrations and in the presence of serum components. Taking these issues into consideration, the heparin detection scheme offered for serum samples cannot be reproduced.

Natural DNA assisted white light generation and stimuli responsive colour tuning

The unique structure of a natural nucleic acid, calf thymus DNA, which can provide an appropriate scaffold for an efficient cascaded energy transfer among organic chromophores, has been used for the generation of bright and pure white light on UV light excitation. Two most commonly used DNA stains, 4′,6-diamidino-2-phenylindole (DAPI) and ethidium bromide (EB) have been used as a part of the donor-acceptor pairs. We have judiciously selected 10-anthracene-10-yl-3-methylbenzothiazol-3-ium chloride (AnMBTZ), an ultrafast molecular rotor, to act as a bridge between DNA bound DAPI and EB for the cascaded flow of energy. The unique molecular rotor properties of AnMBTZ and its exceptional binding ability with natural DNA help to form a distinct tri-chromophoric system in DNA template which can produce bright and pure white light on UV excitation. Detailed flow of energy from photoexcited DAPI to EB via AnMBTZ has been explored using steady state and time-resolved emission spectroscopy. Further, unique binding nature of AnMBTZ with DNA molecules has been used to modulate the colour of the emission from the present tri-chromophoric system by external stimuli, like salt and temperature. Such unique stimuli responsive multi-chromophoric system in a bio-template has great potential for different lightening applications.

An Ultrafast Molecular-Rotor-Based Fluorescent Turn-On Sensor for the Perrhenate Anion in Aqueous Solution.

Devising sensors for the perrhenate anion in aqueous media is extremely challenging, and has seldom been reported in the literature. Herein, we report a fluorescence turn-on sensor for the perrhenate anion in aqueous media based on the aggregation-induced emission of a popular ultrafast molecular rotor dye, Thioflavin-T. The selective response towards the perrhenate anion has been rationalized in terms of matching water affinity, with the weakly hydrated perrhenate anion spontaneously forming a contact ion pair with the weakly hydrated ultrafast molecule-rotor-based organic cation, Thioflavin-T, which in turn leads to an aggregate assembly that provides a fluorescence turn-on response towards perrhenate. The sensing response of Thioflavin-T has been found to be quite selective towards the perrhenate anion when tested against anions that are ubiquitously present in the environment, such as chloride, nitrate, and sulfate anions. The formation of self-assembled Thioflavin-T aggregates has also been investigated by time-resolved emission and temperature-dependent measurements.

Ratiometric fluorescence turn-on sensing of perrhenate anion, a non-radioactive surrogate of hazardous pertechnetate, in aqueous solution

Fluorescent sensor for the perrhenate/pertechnetate anions in aqueous media are rarely reported in literature as their large size and low charge density poses great challenge to their detection. Herein, we report the first fluorescent turn-on ratiometric sensor for perrhenate anion, exploiting its weak hydration property which leads to an advantageous formation of a contact ion-pair of perrhenate anion with a well-known cationic ultrafast molecular rotor dye, Auramine O. This ion-pair formation, in turn, leads to a fluorescent aggregate assembly that provides an excellent opportunity to detect perrhenate anion in aqueous solution.

Porous crystalline architectures: ultra-fast molecular rotors and dynamics control by gas stimuli

Porous crystalline architectures: ultra-fast molecular rotors and dynamics control by gas stimuli

Photophysics of a molecular rotor inside the block co-polymers

Abstract 4 Auramine O, an ultrafast molecular rotor, a diphenylmethane cationic dye is weakly fluorescent in aqueous media. In this article the excited state twisting dynamics of Auramine O in Pluronic F127 and F108 block co-polymer have been reported by using different types of spectroscopic techniques. We observed interesting ground state as well as excited state phenomenon due to the addition of block co-polymers to the aqueous solution of Auramine O. The fluorescence intensity gradually increases with increase in the concentration of both the block co-polymers along with blue shift in the emission maxima. The average lifetime value of Auramine O enhanced significantly with the addition of both the block co-polymers, respectively. Such changes in the spectral properties of Auramine O in the presence of block co-polymers indicate presence of Auramine O inside the micelle. From fluorescence measurement we have also determined partition coefficient of Auramine O between the micellar environment and the aqueous medium. It was observed that in the case of F127 partition coefficient value is higher compare to F108. We have measured the effect of temperature on the photophysics of Auramine O in the presence of block co-polymers. It was observed that the spectroscopic properties of Auramine O/micelle complex are highly dependent on temperature.

Conformational Control of Ultrafast Molecular Rotor Property: Tuning Viscosity Sensing Efficiency by Twist Angle Variation.

Fluorescent molecular rotors find widespread application in sensing and imaging of microscopic viscosity in complex chemical and biological media. Development of viscosity-sensitive ultrafast molecular rotor (UMR) relies upon the understanding of the excited-state dynamics and their implications for viscosity-dependent fluorescence signaling. Unraveling the structure-property relationship of UMR behavior is of significance toward development of an ultrasensitive fluorescence microviscosity sensor. Herein we show that the ground-state equilibrium conformation has an important role in the ultrafast twisting dynamics of UMRs and consequent viscosity sensing efficiency. Synthesis, photophysics, and ultrafast spectroscopic experiments in conjunction with quantum chemical calculation of a series of UMRs based on dimethylaniline donor and benzimidazolium acceptor with predefined ground-state torsion angle led us to unravel that the ultrafast torsional dynamics around the bond connecting donor and acceptor groups profoundly influences the molecular rotor efficiency. This is the first experimental demonstration of conformational control of small-molecule-based UMR efficiencies which can have wider implication toward development of fluorescence sensors based on the UMR principle. Conformation-controlled UMR efficiency has been shown to exhibit commensurate fluorescence enhancement upon DNA binding.

Frontispiece: Ultrafast Molecular Rotors and Their CO2 Tuning in MOFs with Rod‐Like Ligands

Molecular rotors reorienting in times as short as 0.01 nanoseconds at 150K and endowed with minimal constraints (Ea=0.5 kcal mol-1) were realized in MOFs. This benchmark of motional flexibility, coupled with porosity, entailed unprecedented sensitivity to gases, such as CO2, which could modulate extensively rotor dynamics. Our results highlight how largely crystal dynamics are responsive to the gas environment. For more information see the Communication by A. Comotti, A. Maspero et al. on page 11210 ff.

Ultrafast Molecular Rotors and Their CO2 Tuning in MOFs with Rod-Like Ligands.

A metal organic framework (MOF) engineered to contain in its scaffold rod-like struts featuring ultrafast molecular rotors showed extremely rapid 180 ° flip reorientation with rotational rates of 1011 Hz at 150 K. Crystal-pore accessibility of the MOF allowed the CO2 molecules to enter the cavities and control the rotor spinning speed down to 105 Hz at 150 K. Rotor dynamics, as modulated by CO2 loading/unloading in the porous crystals, was described by proton T1 and 2 H NMR spectroscopy. This strategy enabled the regulation of rotary motion by the diffusion of the gas within the channels and the determination of the energetics of rotary dynamics in the presence of CO2 .

Emissive H-Aggregates of an Ultrafast Molecular Rotor: A Promising Platform for Sensing Heparin.

Constructing "turn on" fluorescent probes for heparin, a most widely used anticoagulant in clinics, from commercially available materials is of great importance, but remains challenging. Here, we report the formation of a rarely observed emissive H-aggregate of an ultrafast molecular rotor dye, Thioflavin-T, in the presence of heparin, which provides an excellent platform for simple, economic and rapid fluorescence turn-on sensing of heparin. Generally, H-aggregates are considered as serious problem in the field of biomolecular sensing, owing to their poorly emissive nature resulting from excitonic interaction. To the best of our knowledge, this is the first report, where contrastingly, the turn-on emission from the H-aggregates has been utilized in the biomolecule sensing scheme, and enables a very efficient and selective detection of a vital biomolecule and a drug with its extensive medical applications, i.e., heparin. Our sensor system offers several advantages including, emission in the biologically advantageous red-region, dual sensing, i.e., both by fluorimetry and colorimetry, and most importantly constructed from in-expensive commercially available dye molecule, which is expected to impart a large impact on the sensing field of heparin. Our system displays good performance in complex biological media of serum samples. The novel Thioflavin-T aggregate emission could be also used to probe the interaction of heparin with its only clinically approved antidote, Protamine.

Evaluation of an Ultrafast Molecular Rotor, Auramine O, as a Fluorescent Amyloid Marker.

Recently, Auramine O (AuO) has been projected as a fluorescent fibril sensor, and it has been claimed that AuO has an advantage over the most extensively utilized fibril marker, Thioflavin-T (ThT), owing to the presence of an additional large red-shifted emission band for AuO, which was observed exclusively for AuO in the presence of fibrillar media and not in protein or buffer media. As fibrils are very rich in β-sheet structure, a fibril sensor should be more specific toward the β-sheet structure so as to produce a large contrast between the fibril form and native protein form, for efficient detection and in vitro mechanistic studies of fibrillation. However, in this report, we show that AuO interacts significantly with the native form of bovine serum albumin (BSA), which is an all-α-helical protein and lacks the β-sheet structure, which are the hallmarks of a fibrillar structure. This strong interaction of AuO with the native form of BSA leads to a large emission enhancement of AuO for the native protein itself, and leads to a low contrast between the BSA protein and its fibrils. More importantly, the large red-shifted emission band of AuO, reported in the presence of human insulin fibrils, and which was projected as its major advantage over ThT, is not observed in the presence of BSA fibrils as well as fibrils from other proteins, such as lysozyme, human serum albumin, and β-lactoglobulin. Thus, our results provide information on the universal applicability of the distinctive and claimed-to-be-advantageous photophysical features reported for AuO in human insulin fibrils towards fibrils from other proteins. Time-resolved fluorescence measurements also support the proposition of a strong interaction of AuO with native BSA. Additionally, tryptophan emission of the protein has been explored to further elucidate the binding mechanism of AuO with native BSA. Evaluation of thermodynamic parameters revealed that the binding of AuO with native BSA involved positive enthalpy and entropy changes, suggesting dominant contributions from hydrophobic and electrostatic interactions toward the association of AuO with native BSA. Molecular docking calculations have been performed to identify the principal binding location of AuO in native BSA.

Interaction of a Julolidine-Based Neutral Ultrafast Molecular Rotor with Natural DNA: Spectroscopic and Molecular Docking Studies.

Ultrafast molecular rotors (UMRs) are reported to be one of the best fluorescent sensors to study different microenvironments, including biomolecules. In the present work, we have explored the possibility of application of a julolidine-based neutral UMR, 9-(2,2-dicyano vinyl) julolidine (DCVJ), as a DNA sensor and studied its mode of binding with DNA in detail using spectroscopic and molecular docking techniques. Our spectroscopic studies indicate that association of DCVJ with DNA leads to a very large enhancement in its emission intensity. Detailed investigation reveals that, despite being a neutral molecule, binding of DCVJ with DNA is largely modulated in the presence of salt. Such an unusual salt effect has been explained by invoking the ion-dipole interaction between DCVJ and the phosphate backbone of DNA. The ion-dipole interaction has also been established by studying the interaction of DCVJ with nucleosides. Detailed time-resolved studies show that the twisting motion around the vinyl bond in DCVJ gets retarded to a great extent because of its association with DNA molecules. Through competitive binding studies, it has also been established that DCVJ also binds to DNA through intercalation. Finally, quantum chemical calculations and molecular docking studies have been performed to confirm the mode of binding of DCVJ with DNA.

PicoGreen: a better amyloid probe than Thioflavin-T.

PicoGreen, a cyanine based ultrafast molecular rotor, shows high affinity towards amyloid fibrils and scores a much better sensitivity than Thioflavin-T, a gold standard probe for amyloid fibrils. Detailed spectroscopic and molecular docking studies have been performed to understand the mode of interaction between PicoGreen and amyloid fibrils.

Ultrafast Torsional Relaxation of Thioflavin-T in Tris(pentafluoroethyl)trifluorophosphate (FAP) Anion-Based Ionic Liquids

Ultrafast spectroscopy on solutes, whose dynamics is very sensitive to the friction in its local environment, has strong potential to report on the microenvironment existing in complex fluids such as ionic liquids. In this work, the torsional relaxation dynamics of Thioflavin-T (ThT), an ultrafast molecular rotor, is investigated in two fluoroalkylphosphate ([FAP])-based ionic liquids, namely, 1-ethyl-3-methylimidazolium tris(pentafluoroethyl)trifluorophosphate ([EMIM][FAP]) and 1-(2-hydroxyethyl)-3-methylimidazolium tris(pentafluoroethyl)trifluorophosphate ([OHEMIM][FAP]), using ultrafast fluorescence up-conversion spectroscopy. The emission quantum yield and the excited-state fluorescence lifetime measurement suggest that the torsional relaxation of Thioflavin-T, in this class of ionic liquids, is guided by the viscosity of the medium. The temporal profile of the dynamic Stokes' shift of ThT, measured from time-resolved emission spectrum (TRES), displays a multiexponential behavior in both ionic liquids. The long time dynamics of the Stokes' shift is reasonably slower for the hydroxyethyl derivative as compared to the ethyl derivative, which is in accordance with their measured shear viscosity. However, the short time dynamics of Stokes' shift is very similar in both the ionic liquids, and seems to be independent of the measured shear viscosity of the ionic liquid. We rationalize these observations in terms of different locations of ThT in these ionic liquids. These results suggest that despite having a higher bulk viscosity in the ionic liquid, they can provide unique microenvironment in their complex structure, where the reaction can be faster than expected from their measured shear viscosity.

An ultrafast molecular rotor based ternary complex in a nanocavity: a potential "turn on" fluorescence sensor for the hydrocarbon chain.

Formation of a ternary complex by an ultrafast molecular rotor (UMR) with a macrocyclic cavitand has been investigated for the sensitive detection of the alkyl chain of a surfactant. A benzothiazole based UMR, Thioflavin-T (ThT), has been used as a fluorescent probe. It is shown that ThT forms a very weak inclusion complex with γ-cyclodextrin (γ-CD) with an association constant of 8.8 M(-1). However, the addition of a small amount of surfactant results in a significant increase in the emission intensity of ThT in γ-CD solution. From detailed steady-state and time-resolved fluorescence measurements and NMR studies, it has been inferred that the addition of the surfactant results in the formation of a ternary complex through the inclusion of its alkyl chain inside the γ-CD nanocavity. In such a ternary complex, the non-radiative torsional motion in ThT is largely prevented due to a large increase in the frictional force inside the nanocavity and results in a significant fluorescence enhancement. The formation of the binary and the ternary complexes in the present system has been further supported by the molecular docking and subsequent molecular dynamics simulation studies. The present result indicates that the inclusion complex with an UMR as a guest could be a potential candidate for the efficient detection of insoluble organic molecules, especially hydrocarbons.

Ultrafast molecular rotor based DNA sensor: An insight into the mode of interaction

Abstract Benzothiazole based ultrafast molecular rotor, Thioflavin T (ThT), is a well known fluorescence sensor for nucleic acids. Thus, the knowledge of mode of interaction between ThT and DNA is very important for efficient utilization of such fluorescence sensors. In the present article, we have reported detail studies on the mode of interaction between ThT and a natural DNA using steady-state and femtosecond time-resolved transient emission techniques. Detail steady-state studies show that ThT binds strongly with natural DNA and causes a large change in its spectroscopic properties. Femtosecond transient emission studies show a substantial decrease in the torsional motion in the excited state of ThT bound to DNA molecules, indicating a strong interaction between them. It is observed that ThT binds to DNA through two modes of interaction. Studies with salt indicate that large fraction of ThT binds to DNA by electrostatic interaction, whereas, viscosity and resonance energy transfer studies indicate the intercalation of ThT with DNA molecules.

Molecular Recognition Controlled Delivery of a Small Molecule from a Nanocarrier to Natural DNA

Controlled and targeted release of an active small molecule at the site of demand is very crucial in pharmaceutical applications. In the present article, we have reported a very simple yet unique chemical system which can be used for the controlled and quantitative transfer of a small molecule from a nanocarrier to natural DNA using an external stimulus. Due to the high sensitivity of emission intensity toward its microenvironments, an ultrafast molecular rotor has been used as a spectroscopic probe. SDS micelle has been used as a nanocarrier and the cyclodextrin molecules are used as an external stimulus. The molecular recognition property of the stimulus toward the hydrophobic chain of the surfactant molecules has been utilized for controlled transfer of the small molecule from the nanocarrier to DNA. Through detailed steady state and time-resolved spectroscopic studies, it has been demonstrated that quantitative transfer of the small molecules from nanocarrier to the natural DNA molecules could be achieved. The present chemical system might be very promising in the field of controlled and targeted drug delivery.

Probing the DNA–ionic liquid interaction using an ultrafast molecular rotor

The interaction of imidazolium based ionic liquids with calf thymus DNA has been investigated using a benzothiazole based ultrafast molecular rotor, Thioflavin-T, as a local reporter. It has been found that the addition of ionic liquid causes large reduction in the emission yield of the DNA bound probe indicating a strong interaction between the ionic liquids and DNA. Detailed studies show that the interaction between ionic liquids and DNA is sufficiently strong to remove the bound probe from the DNA. The excited state lifetime of the DNA bound probe measured by femtosecond fluorescence upconversion technique also corroborate with the above proposition. To understand the nature of the interaction between ionic liquids and DNA, photophysical properties of Thioflavin-T in DNA was investigated in the presence of imidazolium based ionic liquids having alkyl side chain of different lengths in the cationic moiety and the results indicate that apart from electrostatic interaction, the hydrophobicity of ionic liquid also plays a significant role in the ionic liquid–DNA interaction.

Ultrafast molecular rotor: an efficient sensor for premelting of natural DNA

Structural changes in nucleic acids in the premelting region (T < melting temperature, T(m)) play an important role in the biological activity of DNA at physiological temperature. In the present communication we report the use of an ultrafast molecular rotor as an extrinsic fluorescence sensor to monitor the structural changes in natural DNA at T < T(m), which could not be detected even by circular dichroism spectroscopy. Further, the fluorescence sensor used in the present study is superior than most commonly used DNA stains.