Cyanine-based Dyes Research Articles

Complexes of photosensitive surfactant and fluorescent dye for light-induced manipulation of colloids

Light-driven diffusioosmosis is a membrane-free method for manipulating colloidal ensembles at solid–liquid interfaces based on photo-sensitive molecules inducing fluid flows along solid surfaces. In this study, we present our findings on porous colloids settled at a solid wall in an aqueous solution comprising a photo-sensitive azobenzene-containing cationic surfactant and a cyanine-based dye, capable of ionically binding to each other. The surfactant acts as an activation agent for diffusioosmotic flow. When exposed to modulated light, it undergoes photo-isomerization from a hydrophobic trans-state to a more hydrophilic cis-state, creating a concentration gradient near the irradiated area of the wall. The resulting osmotic pressure gradient sets the flow in motion. Porous colloids actively participate in flow generation by readily incorporating the surfactant molecules in the trans-state and releasing them in the cis-state, creating a constant source of diffusioosmotic flow. Under UV illumination, an excess of cis-isomers near the porous colloids elicits long-range repulsive interactions, tenfold the diameter of a particle. The dye acts as a sensor for the surfactant filling or emptying the pores of the colloids. It forms a complex with the trans-isomer and diffuses into the pores, where photoisomerization to cis-state destroys the complex and causes both the dye and the surfactant to leave the pores, altering the luminescence brightness within the colloids. We demonstrated that the presence of the dye affects cis-trans isomer ratios of the surfactant at photo-stationary states, thereby influencing the process of diffusioosmosis. This process enables the manipulation of colloidal particles and remote control of the interaction potential between them, facilitating the formation of well-ordered surface aggregates.

Investigation of cyanine-based infrared dyes as calibrants in radiochromic films.

Radiochromic material such as lithium pentacosa-10,12-diynoate (LiPCDA) has been suggested as the radiation-sensitive material for real-time in vivo fiber-optic dosimetry. In this configuration, micron-thick radiochromic coating would measure the absorbed dose, where a major challenge is the uncertainty in the active material thickness, necessitating calibration. A homogeneously incorporated inert infrared (IR) dye, which must also be stable in ambient conditions and against radiolysis, can be added to the radiochromic film to enable optical calibration. This study investigates four commercial cyanine-based dyes (IR-783, IR-806, IR-868, and IR-880) for use as an optical calibrant in fiber-optic radiochromic dosimeters. All dyes were dissolved in water to confirm solubility. IR-783 and IR-806 were dissolved in 10% w/w gelatin solution and coated onto a polyester substrate, which were then sandwiched between two layers of adhesives forming IR-783 and IR-806 films. A second batch of IR dyes in gelatin incorporated the LiPCDA, and was coated onto substrate and sandwiched between adhesive to form IR dye+LiPCDA films. The absorbance spectra of the films were measured periodically (176 and 102 days for IR-dye films, and IR dye+LiPCDA, respectively). The average percentage absorbance, normalized to day 1, was fit to either a single or a double exponential decay model to calculate the spectral stability lifetime (τ1 , τ2 ). Films were irradiated using a 6 MV LINAC beam with a standard setup of 100 source to axis distance (SAD), 10cm×10cm field size and 1.5cm depth. The change in absorbance of the IR-dye+LiPCDA films were measured after they were irradiated to 1, 2, 5, 10, and 20Gy at 3Gy/min. Only IR-783 and IR-806 were sufficiently water soluble. In gelatin matrix, these dyes demonstrated a decrease in absorbance with time for IR-783 and IR-806 dyes, with IR-783 films having an average τ1 =73±7 days and IR-806 films τ1 =7±3 days. When combined with LiPCDA, IR-806 degraded, losing its original peak at ∼820nm. Similarly, IR-783, combined with LiPCDA, showed signs of degradation; however, its original absorbance peak was still observed at ∼800nm. In the IR-783+LiPCDA films, the IR-783 dye had a τ=4±1 days, an order of magnitude faster than the IR-783 with no LiPCDA films. When exposed to x-ray irradiation, the IR-783 dye in the IR-783+LiPCDA films showed no change in absorbance with increasing absorbed dose. In contrast, the LiPCDA in the films responded as expected, increasing in optical density with increased absorbed dose. IR-783 and IR-806 dyes were observed to degrade over time following exponential decay curves. IR-806 could not be combined with the LiPCDA without degrading. The combination of IR-783 with LiPCDA demonstrated single exponential decay behavior at a comparatively faster rate than films that did not have LiPCDA. IR-783 was insensitive to ionizing radiation and thus may be suitable for thickness correction, but an alternative manufacturing procedure may need to be developed.

Squaraine Dye-Sensitized Upconversion with Enhanced Stability and Minimized Aggregation-Caused Quenching.

Upconversion nanoparticles (UCNPs) doped with lanthanides have limited brightness due to their small absorption cross section to light. However, using organic sensitizers can significantly enhance their light absorption ability. Unfortunately, the practical application of organic sensitizers has been hindered by poor stability and aggregation-caused quenching (ACQ). To address these issues, we developed a novel squaraine-based dye, SQ-739, for sensitizing upconversion luminescence (UCL). This dye has a maximum absorption at 739 nm, and shows 1 order of magnitude and 2-fold improved chemical- and photostability, compared to the commonly used cyanine-based dye IR-806, respectively. When SQ-739 is used to sensitize UCNPs, the resulting SQ-739-UCNPs exhibit excellent photostability and reduced ACQ in the presence of polar solvents. Moreover, at the single particle level, the SQ-739-UCNPs exhibit a 97-fold increase in UCL emission compared to bare UCNPs. This squaraine dye-based system represents a new design strategy for developing highly stable and efficient NIR upconversion probes.

Cyanine-based dyes: highly sensitive sensors for the detection of the hypochlorite levels in drinking water based on fluorometry.

Cyanine-based dyes: highly sensitive sensors for the detection of the hypochlorite levels in drinking water based on fluorometry.

Development of thermally-stable NIR absorbing films based on heptamethine cyanine dyes with bistriflimide anion

Near-infrared (NIR) absorbing film is an essential component of CMOS image sensor modules. In order to exploit the intense NIR absorbing performance of heptamethine cyanine dyes for this application, their thermal stability must be improved. In this study, a series of heptamethine cyanine dyes were synthesized in which the meso-substituent and the counter anion were varied. Furthermore, the effects of the substituent and the counter anion on the material properties were investigated. The molar extinction coefficients of the cyanine dyes were enhanced by replacing the iodide ion with the bulky bistriflimide (TFSI) anion. The TFSI anion, in which the negative charge is dispersed over a large volume, stabilized the cyanine cation by charge delocalization and greatly improved the thermal stability. NIR absorbing films with outstanding transmittance in the visible region were fabricated using the synthesized cyanine dyes and a transparent polysulfone (PSU) binder. The thermal stability was evaluated by post-baking the prepared films at a temperature of 150 °C for 3 h. The films made with TFSI-substituted cyanine dyes showed excellent thermal stability, and they maintained their initial absorbance even after the baking process compared to films made with unsubstituted cyanine-based dyes.

J-aggregates albumin-based NIR-II fluorescent dye nanoparticles for cancer phototheranostics.

Phototheranostics, relying on energy conversions of fluorophores upon excitation, integrating diagnostic fluorescence imaging and photo-driven therapy, represents a promising strategy for cancer precision medicine. Compared with the first near-infrared biological window (NIR-I), fluorophores imaged in the second window (NIR-II, 1000–1700 ​nm) exhibit a higher temporal and spatial resolution and tissue penetration depth. Polymethine cyanine-based dye IR1061 is a typical NIR-II small-molecule organic fluorophore, but its low water solubility and short circulation time limiting its biological applications. Therefore, human serum albumin (HSA) nanoparticles with great biocompatibility and biosafety were employed to fabricate hydrophobic IR1061, which exhibited red-shifted absorption band as typical for J-aggregates. Moreover, IR1061@HSA nanoparticles can be successfully used for NIR-II imaging to noninvasively visualize the tumor vascular networks, as well as real-time intraoperative image-guided tumor resection. Interestingly, benefiting from the high photothermal conversion efficiency brought by J-aggregates, IR1061@HSA nanoparticles were also explored for photothermal therapy (PTT) and cause efficient thermal ablation of tumors. Overall, IR1061@HSA, as a novel J-aggregates albumin-based NIR II dye nanoparticle with high biocompatibility, provides an integrated versatile platform for cancer phototheranostics with promising clinical translation prospects.

Synthesis, photophysical, electrochemical and computational investigation of dimethine and trimethine cyanine-based dyes

Three novel cyanine-based dyes (D1-D3) were obtained by the carboxylic indolium salt the condensation reaction with three different donors, N,N-dimethylaniline, carbazole and indole derivatives. The chemical structures of these dyes were confirmed and fully characterized by IR, NMR, and HRMS. The UV–vis, fluorescence spectra, and cyclic voltammetry were conducted to give a preliminary insight into their optoelectronic properties. Generally, all synthesized dyes exhibited well-defined absorption in the visible region with high absorptivity and low band gap, giving λEx. max (516–564 nm), ε (45279–141011 M−1cm−1) and E0-0 (2.11–2.21 eV). Distinctive stokes shifts were observed for all dyes, with the highest value for the dimethine cyanine containing carbazole donor, D2, (2752 cm−1). The solvatochromic study showed hypsochromic effects and increased Stokes shifts with increasing solvent polarity. The electrochemical study on the other hand revealed having HOMO and LUMO energy levels, <-5.46, and > -3.58 eV, respectively, indicating their suitability as sensitizers in dye sensitized solar cells. The impact of different donors on the geometrical, photophysical, charge transfer and reorganization energy on the designed dyes has been investigated using Density Functional Theory (DFT) and time-dependent (TD-DFT). The theoretical and experimental results are in good agreement. The results indicate that the D1-D3 dyes have a small energy gap (2.50–2.77 eV) and their cationic counterpart absorption in the visible (480–576 nm). The modification of the donor suggests that these dyes would be suitable for dye-sensitized solar cells (DSCCs).

Photoexcitation Processes in Oligomethine Cyanine Dyes for Dye-Sensitized Solar Cells-Synthesis and Computational Study.

We report density functional theory (DFT) calculations of three newly synthesized oligomethine cyanine-based dyes as potential TiO2-sensitizers in dye-sensitized solar cells. The three dyes have π-symmetry and the same acceptor side, terminating in the carboxylic anchor, but they differ through the π-bridge and the donor groups. We perform DFT and time-dependent DFT studies and present the electronic structure and optical properties of the dyes alone as well as adsorbed to the TiO2 nanocluster, to provide some predictions on the photovoltaic performance of the system. We analyze theoretically the factors that can influence the short circuit current and the open circuit voltage of the dye-sensitized solar cells. We examine the matching of the absorption spectra of the dye and dye-nanocluster system with the solar irradiation spectrum. We display the energy level diagrams and discuss the alignment between the excited state of the dyes and the conduction band edge of the oxide as well as between the redox level of the electrolyte and the ground state of the dyes. We determine the electron density of the key molecular orbitals and analyze comparatively the electron transfer from the dye to the semiconducting substrate. To put our findings in the right perspective we compare the results of our calculations with those obtained for a coumarin-based dye used in fabricating and testing actual devices, for which experimental data regarding the photovoltaic performance are available.

DNA-Templated Aggregates of Strongly Coupled Cyanine Dyes: Nonradiative Decay Governs Exciton Lifetimes.

Molecular excitons are used in a variety of applications including light harvesting, optoelectronics, and nanoscale computing. Controlled aggregation via covalent attachment of dyes to DNA templates is a promising aggregate assembly technique that enables the design of extended dye networks. However, there are few studies of exciton dynamics in DNA-templated dye aggregates. We report time-resolved excited-state dynamics measurements of two cyanine-based dye aggregates, a J-like dimer and an H-like tetramer, formed through DNA-templating of covalently attached dyes. Time-resolved fluorescence and transient absorption indicate that nonradiative decay, in the form of internal conversion, dominates the aggregate ground state recovery dynamics, with singlet exciton lifetimes on the order of tens of picoseconds for the aggregates versus nanoseconds for the monomer. These results highlight the importance of circumventing nonradiative decay pathways in the future design of DNA-templated dye aggregates.

Millisecond cellular labelling in situ with two-photon photoconversion.

In situ labeling of cells within living biological tissues using photoconversion has provided valuable information on cellular physiology in their natural environments. However, current photoconvertible probes typically require seconds to minutes of light exposure, limiting their uses in rapid biological processes such as intracellular diffusion and circulating cells. Here, we report that two-photon photoconversion of cyanine-based dyes offers unprecedentedly rapid photoconversion down to millisecond timescales per cell. We demonstrate potential biological applications including measuring intracellular diffusion kinetics in a spinal nerve, labeling of rapidly flowing cells in a microfluidic channel, and photoconversion of a circulating cell in vivo.

Locally Excited State-Charge Transfer State Coupled Dyes as Optically Responsive Neuron Firing Probes.

A selection of NIR-optically responsive neuron probes was produced comprising of a donor julolidyl group connected to a BODIPY core and several different styryl and vinylpyridinyl derived acceptor moieties. The strength of the donor-acceptor interaction was systematically modulated by altering the electron withdrawing nature of the aryl unit. The fluorescence quantum yield was observed to decrease as the electron withdrawing effect of the aryl subunit increased in line with changes of the Hammett parameter. The effectiveness of these fluorophores as optically responsive dyes for neuronal imaging was assessed by measuring the toxicity and signal-to-noise ratio (SNR) of each dye. A great improvement of SNR was obtained when compared to the first-generation BODIPY-based voltage sensitive dyes with concomitant toxicity decrease. The mechanism for the optical response is disparate from conventional cyanine-based dyes, opening up a new way to produce effective voltage sensitive dyes that respond well into the NIR region.

Two-photon excited photoconversion of cyanine-based dyes.

The advent of phototransformable fluorescent proteins has led to significant advances in optical imaging, including the unambiguous tracking of cells over large spatiotemporal scales. However, these proteins typically require activating light in the UV-blue spectrum, which limits their in vivo applicability due to poor light penetration and associated phototoxicity on cells and tissue. We report that cyanine-based, organic dyes can be efficiently photoconverted by nonlinear excitation at the near infrared (NIR) window. Photoconversion likely involves singlet-oxygen mediated photochemical cleavage, yielding blue-shifted fluorescent products. Using SYTO62, a biocompatible and cell-permeable dye, we demonstrate photoconversion in a variety of cell lines, including depth-resolved labeling of cells in 3D culture. Two-photon photoconversion of cyanine-based dyes offer several advantages over existing photoconvertible proteins, including use of minimally toxic NIR light, labeling without need for genetic intervention, rapid kinetics, remote subsurface targeting, and long persistence of photoconverted signal. These findings are expected to be useful for applications involving rapid labeling of cells deep in tissue.

Recent Progress in Light-Triggered Nanotheranostics for Cancer Treatment.

Treatments of high specificity are desirable for cancer therapy. Light-triggered nanotheranostics (LTN) mediated cancer therapy could be one such treatment, as they make it possible to visualize and treat the tumor specifically in a light-controlled manner with a single injection. Because of their great potential in cancer therapy, many novel and powerful LTNs have been developed, and are mainly prepared from photosensitizers (PSs) ranging from small organic dyes such as porphyrin- and cyanine-based dyes, semiconducting polymers, to inorganic nanomaterials such as gold nanoparticles, transition metal chalcogenides, carbon nanotubes and graphene. Using LTNs and localized irradiation in combination, complete tumor ablation could be achieved in tumor-bearing animal models without causing significant toxicity. Given their great advances and promising future, we herein review LTNs that have been tested in vivo with a highlight on progress that has been made in the past a couple of years. The current challenges faced by these LTNs are also briefly discussed.

Taking up the cyanine challenge with quantum tools.

ConspectusCyanine derivatives, named from the Greek word kyanos meaning dark-blue, were discovered more than 150 years ago and remain one of the most widely used classes of organic dyes with contemporary applications in photography (panchromatic emulsions), information storage (CD-R and DVD-R media) and biochemistry (DNA and protein labeling) fields. Cyanine chromogens consist of a charged π-conjugated segment containing an odd number of sp2 carbon atoms with the chain capped at the extremities by two electronegative centers, typically nitrogen or oxygen atoms. Cyanines are characterized by a vanishing bond length alternation indicating nearly equal carbon–carbon bond lengths, as well as a very intense and sharp absorption band presenting a shoulder. This hallmark band undergoes a strong red shift when the chain is extended. This so-called vinyl shift is extremely large (ca. 100 nm for each pair of carbon atoms added in the π-conjugated path), making cyanines ideal building blocks for the design of devices with near-infrared applications. Numerous cyanines also exhibit emission bands with large quantum yields. These exceptional optical properties explain why both canonical cyanines and the corresponding fluoroborates (e.g., boron-dipyrromethene, BODIPY) remain the focus of an ever-growing body of experimental work.In turn, this popularity has stimulated quantum mechanical investigations aiming, on the one hand, at probing the specific electronic nature of cyanine dyes and, on the other hand, at helping to design new dyes. However, the adiabatic approximation to time-dependent density functional theory, the most widespread ab initio model for electronically excited states, fails to accurately reproduce the absorption spectra of cyanine derivatives: it yields a systematic and large underestimation of the experimental wavelengths irrespective of the details of the computational protocol. In contrast, highly correlated wave function approaches provide accurate transition energies for model systems but are hardly applicable to real-life cyanines and BODIPY. This indicates that setting up a computationally tractable theoretical protocol that provides both robust and accurate optical spectra for cyanine-based dyes is a major challenge that has only been taken up lately.In this Account, we compile the most recent advances in the field by considering both compact streptocyanines and large fluoroborates. For the former, we summarize the key results obtained with a large panel of theoretical approaches, allowing us not only to understand the origin of the cyanine challenge but also to pinpoint the schemes presenting the most promising accuracy/effort ratio. For the latter, we show via selected examples how theoretical models can be used to reproduce simultaneously experimental band shapes and transition energies, thus paving the way to an efficient in silico design of new compounds.

Activatable Organic Near-Infrared Fluorescent Probes Based on a Bacteriochlorin Platform: Synthesis and Multicolor in Vivo Imaging with a Single Excitation

Near infrared (NIR) fluorescent probes are ideal for in vivo imaging because they offer deeper tissue penetration and lower background autofluorescence. Although most fluorophores in this range are cyanine-based dyes, several new classes of fluorescent NIR probes have been developed. In this study, we developed organic bacteriochlorin derivatives, NMP4 and NMP5, which are excited with a single green light and emit different narrow, well-resolved bands in the NIR (peak of 739 and 770 nm for NMP4 and NMP5, respectively). When conjugated to galactosyl-human serum albumin (hGSA) or glucosyl-human serum albumin (glu-HSA), both targeting H-type lectins, including the β-d-galactose receptor expressing on ovarian cancer, these agents become targeted, activatable, single excitation, multicolor NIR fluorescence probes. After conjugation to either glu-HSA or hGSA, substantial quenching of fluorescence occurs that is reversed after cell binding and internalization. In vitro studies showed higher cancer cell uptake with NMP4 or NMP5 conjugated to hGSA compared to the same conjugates with glu-HSA. In vivo single excitation two-color imaging was performed after intraperitoneal injection of these agents into mice with disseminated ovarian cancer. Excited with a single green light, distinct NIR emission spectra from each fluorophore were detected and could be distinguished with spectral unmixing. In vivo results using a red fluorescence protein (RFP) labeled tumor model of disseminated ovarian cancer demonstrated high sensitivity and specificity for all probes. The success of single excitation, 2-color NIR fluorescence imaging with a new class of bacteriochlorin-based activatable fluorophores, NMP4 and NMP5, paves the way for further exploration of noncyanine dye-based NIR fluorophores.

Practical Labeling Methodology for Choline‐Derived Lipids and Applications in Live Cell Fluorescence Imaging

Lipids of the plasma membrane participate in a variety of biological processes, and methods to probe their function and cellular location are essential to understanding biochemical mechanisms. Previous reports have established that phosphocholine-containing lipids can be labeled by alkyne groups through metabolic incorporation. Herein, we have tested alkyne, azide and ketone-containing derivatives of choline as metabolic labels of choline-containing lipids in cells. We also show that 17-octadecynoic acid can be used as a complementary metabolic label for lipid acyl chains. We provide methods for the synthesis of cyanine-based dyes that are reactive with alkyne, azide and ketone metabolic labels. Using an improved method for fluorophore conjugation to azide or alkyne-modified lipids by Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC), we apply this methodology in cells. Lipid-labeled cell membranes were then interrogated using flow cytometry and fluorescence microscopy. Furthermore, we explored the utility of this labeling strategy for use in live cell experiments. We demonstrate measurements of lipid dynamics (lateral mobility) by fluorescence photobleaching recovery (FPR). In addition, we show that adhesion of cells to specific surfaces can be accomplished by chemically linking membrane lipids to a functionalized surface. The strategies described provide robust methods for introducing bioorthogonal labels into native lipids.

New fluorogenic dyes for analysis of cellular processes by flow cytometry and confocal microscopy

Fluorescent microscopy and fluorescent imaging by flow cytometry are two of the fastest growing areas in the medical and biological research. Innovations in fluorescent chemistry and synthesis of new dye probes are closely related to the development of service equipment such as light sources, and detection techniques. Among compounds known as fluorescent labels, the cyanine-based dyes have become widely used since they have high excitation coefficients, narrow emission bands and high fluorescence upon binding to nucleic acids.The key methods for evaluation of apoptosis and cell cycle allow measuring DNA content by several flow cytometric techniques.We have synthesized new monomethine cyanine dyes and have characterized their applicability for staining of live and/or apoptotic cells. Imaging experiments by flow cytometry and confocal laser scanning microscopy (CLSM) have been also performed. Two of the dyes have shown high-affinity binding to the nuclei at high dilutions, up to 10−9M. Flow cytometry and CLSM have confirmed that these dyes labeled selectively non-living, e.g. ethanol-fixed cells that makes them appropriate for estimations of cell viability and apoptosis. The novel structures proved to be appropriate also for analysis of the cell cycle.

Porous silicon as efficient surface enhanced Raman scattering (SERS) substrate

Silver nanocrystallites are obtained through immersion of porous silicon samples in AgNO 3 solutions and a successive thermal annealing. The efficiency of nanostructures as surface enhanced Raman scattering (SERS) substrates is checked on cyanine-based dyes and horseradish peroxidase, evidencing detectable concentrations as low as 10 −7 to 10 −8 M. The substrate efficiency is strictly related to the Ag particle morphology, which could yield to either local surface plasmons (LSP) coupled to individual particles or to inter-particle short-range interaction.

Solid-Phase Methods for the Synthesis of Cyanine Dyes

We report here a series of studies that explore solid-phase methodologies for the synthesis of various cyanine dyes. The scope of the previously reported catch-and-release method using sulfonyl chloride resin(1) has now been extended to include pentamethine and water-soluble cyanine dyes. We also report a new and chemically distinct synthetic strategy, employing the stepwise attack of heterocyclic carbon nucleophiles on immobilized polyene-chain precursors, allowing the clean synthesis of hydrophobic and hydrophilic trimethine and pentamethine dyes from more easily obtained starting materials. Overall, both approaches appear to be robust and versatile strategies to delivering a wide range of cyanine-based dyes in high purity.