Degenerate 2PA Research Articles

Non-Degenerate Two-Photon Absorption of Fluorescent Protein Chromophores.

Two-photon absorption (2PA), where a pair of photons are absorbed simultaneously, is recognized as a potent bioimaging technique, which depends on the quantified 2PA probability, defined as cross-section (σ2PA). The absorbed photons either have equivalent (ω1 = ω2) or different frequencies (ω1 ≠ ω2), where the former is degenerate 2PA (D-2PA) and the latter is nondegenerate 2PA (ND-2PA). ND-2PA is of particular interest since it is a promising imaging technology with flexibility of photon frequencies and enhanced cross sections, however, it remains a relatively unexplored area compared to D-2PA. This work utilizes time-dependent density functional theory (TD-DFT) and second-order approximate coupled-cluster with the resolution-of-identity approximation (RI-CC2), for the excitation from S0 to S1, to investigate σD-2PA and σND-2PA of FP chromophore models. Interestingly, comparing CAM-B3LYP with the RI-CC2 computations shows qualitative and, in fact, near quantitative agreement in the computed improvements of σND-2PA for comparable (relative) frequency detunings, despite the known underestimations of 2PA cross sections, for TD-DFT results relative to RI-CC2 values. As expected from the 2-state model, the computed values of σND-2PA are quantitatively larger than σD-2PA, where chromophores with the largest values of σD-2PA show greater potential for σND-2PA improvement. Anionic chromophores demonstrated improvements up to 14%, while substantial enhancements were observed in neutral chromophores with some achieving a 30% increase. This work investigates the ND-2PA photophysical characteristics of FP chromophores and identifies qualitative patterns in the computed properties of ND-2PA relative to D-2PA.

Nature of Linear Spectral Properties and Fast Relaxations in the Excited States and Two-Photon Absorption Efficiency of 3-Thiazolyl and 3-Phenyltiazolyl Coumarin Derivatives.

Coumarin-based fluorescent agents play an important role in the manifold fundamental scientific and technological areas and need to be carefully studied. In this research, linear photophysics, photochemistry, fast vibronic relaxations, and two-photon absorption (2PA) of the coumarin derivatives, methyl 4-[2-(7-methoxy-2-oxo-chromen-3-yl)thiazol-4-yl]butanoate (1) and methyl 4-[4-[2-(7-methoxy-2-oxo-chromen-3-yl)thiazol-4-yl]phenoxy]butanoate (2), were comprehensively analyzed using stationary and time-resolved spectroscopic techniques, along with quantum-chemical calculations. The steady-state one-photon absorption, fluorescence emission, and excitation anisotropy spectra, as well as 3D fluorescence maps of 3-hetarylcoumarins 1 and 2 were obtained at room temperature in solvents of different polarities. The nature of relatively large Stokes shifts (∼4000-6000 cm-1), specific solvatochromic behavior, weak electronic π → π* transitions, and adherence to Kasha's rule were revealed. The photochemical stability of 1 and 2 was explored quantitatively, and values of photodecomposition quantum yields, on the order of ∼10-4, were determined. A femtosecond transient absorption pump-probe technique was used for the investigation of fast vibronic relaxation and excited-state absorption processes in 1 and 2, while the possibility of efficient optical gain was shown for 1 in acetonitrile. The degenerate 2PA spectra of 1 and 2 were measured by an open aperture z-scan method, and the maximum 2PA cross-sections of ∼300 GM were obtained. The electronic nature of the hetaryl coumarins was analyzed by quantum-chemical calculations using DFT/TD-DFT level of theory and was found to be in good agreement with experimental data.

New Two-Photon Absorbing Squaraine Derivative with Efficient Near-Infrared Fluorescence, Superluminescence, and High Photostability

The nature of linear photophysical and nonlinear optical properties of a new squaraine derivative 2,4-bis[4-(azetidyl)-2-hydroxyphenyl]squaraine (1) with efficient near-infrared (NIR) emission was comprehensively analyzed based on spectroscopic, photochemical, and two-photon absorption (2PA) measurements, along with quantum chemical analysis. The steady-state absorption, fluorescence, and excitation anisotropy spectra of 1 and its fluorescence emission lifetimes revealed the multiple aspects of the electronic structure of 1, including the relative orientations of the main transition dipoles, effective rotational volumes in solvents of different polarities, and a maximum molar extinction of 1.35 × 10-5 M-1·cm-1, which is unusually small for similar symmetric squaraines. The degenerate 2PA spectrum of 1 was obtained over a broad spectral range under femtosecond excitation, using standard open-aperture Z-scan and two-photon induced fluorescence methods, revealing maximum 2PA cross sections of ∼400 GM. Squaraine 1 exhibited efficient superluminescence emission in the polar solvent (dichloromethane) at room temperature under femtosecond pumping conditions. Quantum chemical analysis of the electronic structure of 1 was performed using the DFT/TD-DFT level of theory and found to be in good agreement with experimental data. The new squaraine derivative 1 displayed high fluorescence quantum yield, efficient NIR superluminescence, large 2PA cross sections, and high photostability with a photodecomposition quantum yield ∼4 × 10-6, suggesting its potential for applications in two-photon fluorescent bioimaging and lasing.

Enhanced two-photon absorption of sandwich-like coordination complexes based on squaraine and metallomacrocycle derivatives

Covalent or noncovalent multi-chromophore structures have expanded the discovery of appreciable nonlinear optical (NLO) materials. We present here a report of how drastic changes in two-photon absorption (2PA) coefficients can enhance from slight changes of electronic delocalization through coordination interaction. Experimental results illustrate that 2PA is the dominant phenomenon contributing to the NLO properties of the complexes recorded in the range of 750–1550 nm by open aperture Z-scan method. The values of degenerate 2PA cross sections (δ2PA) can be enhanced obviously by coordination interactions assisted by steric effect and intermolecular charge transfer, not showing a simply sum of the nonlinear parameters of the starting components. Importantly, the complexes feature high δ2PA values up to 12,700 GM at 1400 nm in the second near-infrared region. Both complexes with large 2PA coefficients and multiple excitation wavelengths may offer a novel avenue for creating useful nonlinear absorbing materials.

Electronic Nature of Neutral and Charged Two-Photon Absorbing Squaraines for Fluorescence Bioimaging Application

The electronic properties of neutral 2,4-bis(4-bis(2-hydroxyethyl) amino-2-hydroxy-6-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)phenyl)squaraine (1) and charged 2-((3-octadecylbenzothiazol-2(3H)-ylidene)methyl)-3-oxo-4-((3-(4-(pyridinium-1-yl)butyl)benzo-thiazol-3-ium-2-yl)methylene)cyclobut-1-enolate iodide (2) squaraine derivatives were analyzed based on comprehensive linear photophysical, photochemical, nonlinear optical studies (including two-photon absorption (2PA) and femtosecond transient absorption spectroscopy measurements), and quantum chemical calculations. The steady-state absorption, fluorescence, and excitation anisotropy spectra of these new squaraines revealed the values and mutual orientations of the main transition dipoles of 1 and 2 in solvents of different polarity, while their role in specific nonlinear optical properties was shown. The degenerate 2PA spectra of 1 and 2 exhibited similar shapes, with maximum cross sections of ∼300–400 GM, which were determined by the open aperture Z-scan method over a broad spectral range. The nature of the time-resolved excited-state absorption spectra of 1 and 2 was analyzed using a femtosecond transient absorption pump–probe technique and the characteristic relaxation times of 4–5 ps were revealed. Quantum chemical analyses of the electronic properties of 1 and 2 were performed using the ZINDO/S//DFT theory level, affording good agreement with experimental data. To demonstrate the potential of squaraines 1 and 2 as fluorescent probes for bioimaging, laser scanning fluorescence microscopy images of HeLa cells incubated with new squaraines were obtained.

Dispersive white light continuum single Z-scan for rapid determination of degenerate two-photon absorption spectra

We present an experimental technique to determine the degenerate two-photon absorption (2PA) spectra by performing a single Z-scan using a high-spectral-irradiance white light continuum (WLC) generated by a hollow core fiber. The hollow fiber was filled with Argon (Ar) gas at a pressure of 0.6 bar and was pumped with 500 mJ, 30 fs, and 800 nm pulses. The broadband WLC pulses with 350 nm bandwidth in the range of 600–950 nm were compressed to sub-8 fs pulses. To characterize and interpret the data obtained from this method, the spectral, temporal and spatial characteristics of the WLC were first analyzed. The WLC emerging from the compressor was dispersed using a prism pair and then focused into the sample by a cylindrical lens. Since different spectral components are spatially separated, any part of the sample in the beam cross section is irradiated with almost single wavelength pulses leading to only a degenerate 2PA process. The nonlinear transmittance was then measured by a charge-coupled-device (CCD) line camera as a function of the sample position while the sample was moved along the beam direction by a motorized translation stage. In this way the Z-scans at different wavelengths in the WLC spectral range can be measured and thus the wavelength-resolved degenerate 2PA spectra can be obtained by performing a single scan using dispersive WLC. This method was verified on a well-characterized dye Rhodamine B and yield a reasonable agreement with the data found in the literature. We used this method to determine the 2PA spectra of some two-photon initiators (2PIs) developed for two-photon polymerization (2PP) based 3D micro-structuring.

Electronic Nature of New Ir(III) Complexes: Linear Spectroscopic and Nonlinear Optical Properties

Comprehensive investigations of the linear and nonlinear optical properties of new Ir(III)-complexes [Ir(pbt)2(dbm)] (1), [Ir(pbt)2(dmac)] (2), and [Ir(pbt)2(minc)] (3) (pbt=2-phenylbenzothiazole; dbm=dibenzoyl methane; dmac=(1E,4Z,6E)-1,7-bis(4-(dimethylamino)phenyl)-5-hydroxyhepta-1,4,6-trien-3-one; minc=(1E,4Z,6E)-5-hydroxy-1,7-bis(1-methyl-1H-indol-3-yl)hepta-1,4,6-trien-3-one) are reported, including, photostability, two-photon absorption, and femtosecond transient absorption spectroscopy. The steady-state and time-resolved spectral properties of 1-3 revealed the electronic nature of the absorption bands, and photoluminescence emission of 2 and 3 shows both fluorescence and phosphorescence processes occurring simultaneously in liquid solution at room temperature. This unusual behavior of 2 and 3 can be explained by a dual-minimum potential surface of the excited electronic state resulting in two independent fluorescence and phosphorescence emission channels. The degenerate 2PA spectra of 1-3 were obt...

Linear Photophysics and Femtosecond Nonlinear Spectroscopy of a Star-Shaped Squaraine Derivative with Efficient Two-Photon Absorption

The synthesis, comprehensive linear photophysical and photochemical study, two-photon absorption (2PA) spectrum, ultrafast relaxation kinetics in the excited states, and efficient superluminescence properties of a new symmetrical three-armed star-shaped squaraine derivative (1) are presented. The steady-state spectral parameters of 1 in a number of organic solvents, including fluorescence excitation anisotropy spectra, revealed a weak interaction between the squaraine branches and the effect of symmetry breaking in the ground electronic state. The degenerate 2PA spectrum of 1 was obtained over a broad spectral range with a maximum cross section of ∼8000 GM using the open aperture Z-scan technique. The nature of the fast dynamic processes in the excited electronic states of 1 was investigated by the femtosecond transient absorption pump–probe method, revealing characteristic relaxation times of ∼3–4 ps. The efficient superluminescence emission of 1 was observed in relatively low concentration solution (≈ 2...

Enhancement of Two-Photon Absorption in Quantum Wells for Extremely Nondegenerate Photon Pairs

We recently demonstrated orders of magnitude enhancement of two-photon absorption (2PA) in direct gap semiconductors due to intermediate state resonance enhancement for photons of very different energies. It can be expected that further enhancement of nondegenerate 2PA will be observed in quantum wells (QWs) since the intraband matrix elements do not vanish near the band center as they do in the bulk, and the density of states in QWs is larger near the band edge. Here we present a perturbation-theory based theoretical description of nondegenerate 2PA in semiconductor QWs, where both frequency and polarization of two incident waves can vary independently. Analytical expressions for all possible permutations of frequencies and polarizations have been obtained, and the results are compared with degenerate 2PA in quantum wells along with degenerate and nondegenerate 2PA in bulk semiconductors. We show that using QWs in place of bulk semiconductors with both beams in the TM-polarized mode leads to an additional order of magnitude increase in the nondegenerate 2PA. Explicit calculations for GaAs QWs are presented.

New Fluorene-Based Fluorescent Probe with Efficient Two-Photon Absorption

The synthesis, linear photophysical characterization, and two-photon absorption (2PA) properties of new fluorene derivative 3,30-(pyridine-2,6-diyl)bis(1-(7-(diphenylamino)-9,9-dihexyl-9H-fluoren-2-yl)propane-1,3-dione) (1) have been presented. The steady-state absorption, fluorescence and excitation anisotropy spectra along with the fluorescence decay kinetics of 1 are obtained in the solvents of different polarities at room temperature with respect to its potential application in bioimaging. The analysis of linear photophysical properties revealed a complicated nature of the main one-photon absorption band of 1, and the strong solvatochromic effect in steady-state fluorescence spectra is observed. The degenerate 2PA spectrum of 1 is measured in the spectral range 570–970 nm with the use of the open aperture Z-scan method under the 1-kHz femtosecond excitation, and the maximum values of two-photon action cross sections ~(100–130) GM are obtained. The nature of the linear absorption and the 2PA bands is analyzed by quantum chemical methods using a Gaussian program package.

Two-photon absorption and time-resolved stimulated emission depletion spectroscopy of a new fluorenyl derivative.

The synthesis, comprehensive linear photophysical characterization, two-photon absorption (2PA), steady-state and time-resolved stimulated emission depletion properties of a new fluorene derivative, (E)-1-(2-(di-p-tolylamino)-9,9-diethyl-9H-fluoren-7-yl)-3-(thiophen-2-yl)prop-2-en-1-one (1), are reported. The primary linear spectral properties, including excitation anisotropy, fluorescence lifetimes, and photostability, were investigated in a number of aprotic solvents at room temperature. The degenerate 2PA spectra of 1 were obtained with open-aperture Z-scan and two-photon induced fluorescence methods, using a 1 kHz femtosecond laser system, and maximum 2PA cross-sections of ∼400-600 GM were obtained. The nature of the electronic absorption processes in 1 was investigated by DFT-based quantum chemical methods implemented in the Gaussian 09 program. The one- and two-photon stimulated emission spectra of 1 were measured over a broad spectral range using a femtosecond pump-probe-based fluorescence quenching technique, while a new methodology for time-resolved fluorescence emission spectroscopy is proposed. An effective application of 1 in fluorescence bioimaging was demonstrated by means of one- and two-photon fluorescence microscopy images of HCT 116 cells containing dye encapsulated micelles.

Extremely nondegenerate two-photon absorption in direct-gap semiconductors [Invited

Two-photon absorption (2PA) spectra with pairs of extremely nondegenerate photons are measured in several direct-gap semiconductors (GaAs, CdTe, ZnO, ZnS and ZnSe) using picosecond or femtosecond pulses. In ZnSe, using photons with a ratio of energies of ~12, we obtain a 270-fold enhancement of 2PA when comparing to the corresponding degenerate 2PA coefficient at the average photon energy (ηω1 + ηω2)/2. This corresponds to a pump photon energy of 8% of the bandgap. 2PA coefficients as large as 1 cm/MW are measured. Thus, by using two widely different wavelengths we are able to access the large 2PA observed previously only in narrow gap semiconductors. We also calculate the corresponding enhancement of nonlinear refraction, consisting of two-photon, AC-Stark and Raman contributions. The net effect is a smaller enhancement, but exhibits very large dispersion within the 2PA regime.

A superfluorescent fluorenyl probe with efficient two-photon absorption

The linear photophysical, excited state absorption (ESA), superfluorescence, and two-photon absorption (2PA) properties of 4,4'-(1E,1'E)-2,2'-(7,7'(1E,1'E)2,2'(4,4'-sulfonylbis(4,1-phenylene))bis(ethane-2,1-diyl)bis(9,9-didecy-9H-fluorene7,2-diyl))bis(ethane-2,1-diyl)bis(N,N-diphenylaniline) (1) were investigated in organic and aqueous media with respect to its potential application in biological imaging. The analysis of linear photophysical properties revealed a rather complex nature of the main one-photon absorption band, strong solvatochromic effects in the steady-state fluorescence spectra, single-exponential fluorescence decay, and high fluorescence quantum yields in organic solvents (≈1.0). The ESA spectra of 1 suggested potential for light amplification in nonpolar media while efficient superfluorescence in cyclohexane was demonstrated. The degenerate 2PA spectra of 1 were obtained over a broad spectral range (640-900 nm), using a standard two-photon induced fluorescence method under 1 kHz femtosecond excitation. Two well defined 2PA bands with maximum 2PA cross sections up to 1700 GM in the higher energy, short wavelength band and ≈1200 GM in the lower energy, long wavelength band of 1 were shown. The potential use of 1 in bioimaging was demonstrated via one- and two-photon in vitro fluorescence imaging of HCT 116 cells.

Fluorene-Based Metal-Ion Sensing Probe with High Sensitivity to Zn2+ and Efficient Two-Photon Absorption

The photophysical, photochemical, two-photon absorption (2PA) and metal ion sensing properties of a new fluorene derivative (E)-1-(7-(4-(benzo[d]thiazol-2-yl)styryl)-9,9-bis(2-(2-ethoxyethoxy)ethyl)-9H-fluoren-2-yl)-3-(2-(9,10,16,17,18,19,21,22,23,24-decahydro-6H dibenzo[h,s][1,4,7,11,14,17]trioxatriazacycloicosin-20(7H)-yl)ethyl)thiourea (1) were investigated in organic and aqueous media. High sensitivity and selectivity of 1 to Zn(2+) in tetrahydrofuran and a water/acetonitrile mixture were shown by both absorption and fluorescence titration. The observed complexation processes corresponded to 1:1 stoichiometry with the range of binding constants approximately (2-3) x 10(5) M(-1). The degenerate 2PA spectra of 1 and 1/Zn(2+) complex were obtained in the 640-900 nm spectral range with the maximum values of two-photon action cross section for ligand/metal complex approximately (90-130) GM, using a standard two-photon induced fluorescence methodology under femtosecond excitation. The nature of the 2PA bands was analyzed by quantum chemical methods and a specific dependence on metal ion binding processes was shown. Ratiometric fluorescence detection (420/650 nm) provided a good dynamic range (10(-4) to 10(-6) M) for detecting Zn(2+), which along with the good photostability and 2PA properties of probe 1 makes it a good candidate in two-photon fluorescence microscopy imaging and sensing of Zn ions.

Comparison of nonlinear absorption in three similar dyes: Polymethine, squaraine and tetraone

We performed a detailed experimental investigation and quantum-chemical analysis of two-photon absorption (2PA) spectra of symmetrical cationic polymethine and neutral squaraine and tetraone dyes having nearly identical terminal groups and similar conjugation length. Squaraine and tetraone structures differ from polymethines in that their conjugated backbone includes an electron accepting bridge. Frequency degenerate 2PA spectra of these molecules are measured by two-photon fluorescence spectroscopy and the Z-scan technique. Comparing 2PA spectra of polymethine with squaraine and tetraone structures, we find that we can access considerably larger 2PA cross-sections (⩾8000 × 10 −50 cm 4 s/photon) in the squaraine and tetraone molecules due to an increase in the density of final states introduced by the electron accepting bridge in the conjugated chain. We discuss the difference in electronic structure for these molecules and introduce a link between 2PA and excited-state absorption spectra. The results of these experiments combined with agreement of quantum-chemical calculations moves us closer to a predictive capability for the relation between molecular structure and nonlinear optical properties.

Experimental and theoretical approaches to understanding two-photon absorption spectra in polymethine and squaraine molecules

We performed a detailed experimental investigation and quantum-chemical analysis of two-photon absorption (2PA) spectra of a series of symmetrical cationic polymethines and neutral squaraines having similar structures. Degenerate 2PA spectra of these molecules are taken by using two-photon fluorescence spectroscopy and the Z-scan technique. All measurements are made with 150 fs laser pulses of 1 kHz repetition rate in the tuning range of 520-2100 nm(0.6-2.4 eV). Comparing 2PA spectra of polymethines and squaraines, we find that we can access considerably larger 2PA cross sections (≥8600×10−50 cm4s/photon) in the squaraines owing to narrower linear absorption spectra and an increase in the density of unoccupied molecular orbitals introduced by the squaraine acceptor group in the conjugated chain.

Two-photon absorption in CdTe quantum dots

We report measurements of frequency degenerate and nondegenerate two-photon absorption (2PA) spectra of CdTe quantum dots, QDs, in glass matrices and compare them with 2PA in bulk CdTe. We find that the 2PA is strongly dependent on the size of the QDs becoming smaller with decreasing size, even when normalizing to the volume of the dots. We adapt a simple degenerate 2PA model, based on the effective mass approximation, to nondegenerate 2PA, and this model correctly describes the experimental data for 2-photon energies up to ~ 1.4Eg. This suggests that, once the spectrum for one size of quantum dot is known, the model can be used for predicting the degenerate and nondegenerate 2PA spectra of different sized QDs of the same semiconductor.