Living cells are known to be crowded with organelles, biomembranes, and macromolecules such as proteins, DNA, RNA, and actin filaments. It is believed that such macromolecular crowding affect biomolecular diffusion, protein-protein and protein-substrate interaction, and protein folding. In this contribution, I will discuss our recent results on rotational and translational diffusion of small and large molecules in crowded environments using time-resolved anisotropy and fluorescence correlation spectroscopy methods. In these studies, rhodamine green and enhanced green fluorescent protein are used as fluorescent probes diffusing in buffers enriched with biomimetic crowding agents such as Ficoll-70, bovine serum albumin (BSA), and ovalbumin. Controlled experiments on pure and glycerol-rich buffers were carried out as environments with variable, homogeneous viscosity. Our results indicate that the microviscosity differs from the corresponding bulk viscosity, depending on the nature of crowding agents (i.e., proteins versus polymers), the concentration of crowding agents and spatio-temporal scaling of our experimental approach. Our findings provide a foundation for fluorescence-based studies of diffusion and binding of biomolecules in the crowded milieu of living cells.

We have studied one of the aromatic amino acids, tyrosine, regarding its photophysical properties in various solvent conditions by using a femtosecond fluorescence up-conversion technique and high-level TDDFT and CC2 computations. In this talk, profound details not only on ultrafast solvation dynamics on a neutral tyrosine in various solvents, but also on the excited-state dynamics for a single- (or doubly-) deprotonated tyrosine under various pH solutions will be presented. In high basicity, a tyrosine shows different absorption/emission spectra, and a total spectrum consists of a combination of these individual spectra that depend on the pH of the solution. The time scale of acid-base equilibrium is essential in solvation dynamics; whereas the protonation is simply controlled by diffusion, the de-protonation is considered to be slow process such that acid-base equilibrium may not be reached in the short-lived excited state after photo-excitation. Experimental and computational approaches taken and insights obtained in this concerted work will be described.

Following ultraviolet excitation, the canonical purine nucleobases, guanine and adenine, are able to efficiently dissipate the absorbed energy within hundreds of femtoseconds. This property affords these nucleobases with great photostability. Conversely, non-canonical purine nucleobases exhibit high fluorescence quantum yields or efficiently populate long-lived triplet excited states from which chemistry can occur. Using femtosecond broadband transient absorption spectroscopy in combination with ab initio static and surface hopping dynamics simulations we have determined the electronic and structural factors that regulate the excited state dynamics of the purine nucleobase derivatives. Importantly, we have uncovered that the photostability of the guanine and adenine nucleobases is not due to the structure of the purine core itself and that the substituent at the C6 position of the purine nucleobase plays a more important role than that at the C2 position in the ultrafast relaxation of deleterious electronic energy. [The authors acknowledge the CAREER program of the National Science Foundation (Grant No. CHE-1255084) for financial support.]

The ultrafast dynamics of nucleic acids have been under scrutiny for the past couple of decades because of the role that the high-energy electronic states play in mutagenesis and carcinogenesis. Kinetic models have been proposed, based on both experimental and theoretical discoveries. Direct experimental evidence of the intersystem crossing rate and population of the triplet state for most nucleic acid bases has yet to be reported, even though the triplet state is thought to be the most reactive species. Utilizing broadband femtosecond transient absorption spectroscopy, we reveal the time scale at which singlet-to-triplet population transfer occurs in several nucleic acid derivatives in the condensed phase. The implication of these results to the current understanding of the DNA and RNA photochemistry will be discussed. The authors acknowledge the CAREER program of the National Science Foundation (Grant No. CHE-1255084) for financial support.

The utility of a perspective donor or acceptor molecule for photoelectric applications is difficult to predict a priori. This hinders productive synthetic exploration and necessitates lengthy device optimization procedures for reasonable estimation of said molecule’s applicability. Using femtosecond broadband transient absorption spectroscopy, supported by time-dependent density functional theory computations and steady-state-absorption and emission spectroscopies, we have characterized a family of perspective optoelectronic compounds, in an effort to predict their relative performance in organic photovoltaic devices from information accrued from excited-state dynamics and photophysical properties. A series of tetraphenylazadipyrromethene (ADP) complexes chelated with three different metal centers was investigated. We have determined that the chelating metal has little effect on the ground state properties of this family. However their excited state dynamics are strongly modulated by the metal. Specifically, the zinc-chelated ADP complex remains in the excited state tenfold longer than the cobalt or nickel complexes. We assert that this is key photophysical property that should make the zinc complex outperform the other two complexes in photovoltaic applications. This hypothesis is supported by preliminary power conversion efficiency results in devices.

Semiconductor nanoclusters (SCNCs) are promising electronic materials for use in solid-state device fabrication, where device efficiency is strongly controlled by charge generation and transfer from SCNCs to their surroundings. In this paper we report the excited-state dynamics of molecule-like 1.6 nm diameter CdSe SCNCs, which are passivated with the highly conjugated ligand phenyldithiocarbamate (PDTC) or para-substituted PDTCs. Femtosecond transient absorption studies reveal sub-picosecond hole transfer (τ ≈ 0.9 ps) from a SCNC to its ligand shell based on strong electronic interaction and hole delocalization, and hot electron transfer (τ ≈ 0.2 ps) to interfacial states created by charge separation. A series of control experiments were performed by varying SCNC size (1.6 nm v.s. 2.9 nm) and photon energy of the pump laser (388 nm v.s. 490 nm), as well as addition of electron quencher (benzoquinone) and hole quencher (pyridine), which rules out alternative mechanisms and confirms the critical role of energy level alignment between the SCNC and its passivating ligands.

Two-dimensional pump-probe electronic spectroscopy is a powerful technique able to provide both high spectral and temporal resolution, allowing the analysis of ultrafast complex reactions occurring via complementary pathways by the identification of decay-specific fingerprints. [1-2] The understanding of the origin of the experimentally recorded signals in a two-dimensional electronic spectrum requires the characterization of the electronic states involved in the electronic transitions photoinduced by the pump/probe pulses in the experiment. Such a goal constitutes a considerable computational challenge, since up to 100 states need to be described, for which state-of-the-art methods as RASSCF and RASPT2 have to be wisely employed. [3] With the present contribution, the main features and potentialities of two-dimensional electronic spectroscopy are presented, together with the machinery in continuous development in our groups in order to compute two-dimensional electronic spectra. The results obtained using different level of theory and simulations are shown, bringing as examples the computed two-dimensional electronic spectra for some specific cases studied. [2-4] l0pt Figure [1] Rivalta I, Nenov A, Cerullo G, Mukamel S, Garavelli M, Int. J. Quantum Chem., 2014, 114, 85 [2] Nenov A, Segarra-Martí J, Giussani A, Conti I, Rivalta I, Dumont E, Jaiswal V K, Altavilla S, Mukamel S, Garavelli M, Faraday Discuss. 2015, DOI: 10.1039/C4FD00175C [3] Nenov A, Giussani A, Segarra-Martí J, Jaiswal V K, Rivalta I, Cerullo G, Mukamel S, Garavelli M, J. Chem. Phys. submitted [4] Nenov A, Giussani A, Fingerhut B P, Rivalta I, Dumont E, Mukamel S, Garavelli M, Phys. Chem. Chem. Phys. Submitted [5] Krebs N, Pugliesi I, Hauer J, Riedle E, New J. Phys., 2013,15, 08501

Ultrafast Terahertz Kerr Effect (TKE) spectroscopy is a relatively new nonlinear THz technique that is sensitive to the orientational dynamics of anisotropic, condensed-phase samples. The sample is excited by a single high field strength  ∼ 1 picosecond THz pulse, and the resulting transient birefringence is measured by a  ∼ 40 femtosecond 800 nm probe pulse. We have measured the TKE response of several aromatic liquids at room temperature, including benzene, benzene-d6, hexafluorobenzene, pyridine, and toluene. The measured decay constants range from  ∼ 1-10 ps, and, along with previous optical Kerr effect results in the literatureLoughnane et al. JPCB 110.11 (2006): 5708-5720. give insights into intermolecular interactions in these liquids. Loughnane et al. JPCB 110.11 (2006): 5708-5720.,

The ultrafast TeraHertz Kerr effect (TKE) has recently been demonstrated as a nonlinear spectroscopic technique capable of measuring the dielectric relaxation of liquids. The true power of this technique lies in its ability to provide complementary information to measurements taken using heterodyne-detected optical Kerr effect (OKE) spectroscopy. The optical pulses in OKE measurements interact with the sample via the molecular polarizability, a rank-two tensor, in contrast with THz pulses that interact with the molecules via the dipole moment, a rank-one tensor. Given the different light-matter interactions in the two techniques, TKE measurements help complete the physical picture of intermolecular interactions at short timescales. We report here our implementation of heterodyne-detected TKE spectroscopy, along with measurements of pure liquids, and binary mixtures. Some of the liquids presented here were previously believed to be TKE inactive, thus showing that we have achieved a greater sensitivity than the previous implementation in the literature. In addition, we will discuss a variety of binary mixtures and show how the TKE data can be compared with OKE data to deepen our physical understanding of intermolecular interactions in liquids.

The vibrational and isotopic dependence of the hindered (tunneling-mediated) proton-transfer reaction taking place in the ground electronic state (~X¹A₁) of monodeuterated tropolone (TrOD) has been explored under ambient (bulk-gas) conditions by applying two-color variants of resonant four-wave mixing (RFWM) spectroscopy in conjunction with polarization-resolved detection schemes designed to alleviate spectral complexity and facilitate rovibrational assignments. Full rotation-tunneling analyses of high-resolution spectral profiles acquired for the fundamental and first-overtone bands of a reaction-promoting O−D…O deformation/ring-breathing mode, ν₃₆(a₁), were performed, thereby extracting refined structural and dynamical information that affords benchmarks for the quantitative interpretation of tunneling-induced signatures found in long-range scans of ~X¹A₁ vibrational levels residing below E^X̃_vib = 1700 cm⁻¹. Observed kinetic isotope effects, which reflect changes in both reaction kinematics and vibrational displacements, will be discussed, with high-level quantum-chemical calculations serving to elucidate state-resolved propensities for proton transfer in TrOH and TrOD.

Halomethanes have attracted extensive research efforts of considerable variety, owing to their relative simplicity and ubiquitous presence in synthetic and environmental settings as well as their amenability to benchmark problems in physical chemistry. Their role in atmospheric processes is well known, most famously as the source of atomic halogens which catalyze the depletion of stratospheric ozone. Indeed, the photolytic cleavage of the carbon-halogen bond is the primary fate of halomethanes in the atmosphere. We utilize laser-induced photolysis to study the C-Br bond cleavage in CHBrCl₂ in a molecular beam. Atomic bromine fragments are probed with resonance enhanced multiphoton ionization (REMPI), which allows ground state and spin-excited products to be independently detected. Action spectroscopy in conjunction with velocity map imaging is used to determine the internal energy of the CHCl₂ partner fragment. Product state distributions as a function of photolysis energy may be discerned with these techniques. Current results will be presented.

The lowest energy singlet (ã¹A₁) and triplet (~X³B₁) electronic states of methylene, CH₂, are only separated by 3150 cm⁻¹, but differ greatly in chemical reactivity. Overall methylene reaction rates and chemical behavior are therefore strongly dependent on collisionally-mediated singlet-triplet interconversion. Collisions with inert partners tend to depopulate the excited singlet state and populate vibrationally excited triplet levels in CH₂. This process is generally considered as irreversible for large molecules, however, this is not the case for small molecules such as CH₂. An investigation of the decay kinetics of CH₂ in the presence of argon and various amounts of oxygen has been carried out using transient frequency modulation (FM) absorption spectroscopy, to monitor ortho and para rotational levels in both the ã¹A₁(000) and ã¹A₁(010) states. In the ã¹A₁(000) state, all observed rotational levels follow double exponential decay kinetics, a direct consequence of reversible intersystem crossing. The relative amplitude of the slower decay component is an indicator of how quickly the reverse crossing from excited triplet levels becomes significant during the reaction and relaxation of singlet methylene. The para rotational levels show more obvious signs of reversibility than ortho rotational levels. Adding oxygen enhances the visibility of reversibility for both ortho and para levels. However, in the ã¹A₁(010) state where the FM signal is 5-10 times smaller than the ã¹A₁(000) state, there is no evidence of double exponential decay kinetics. Acknowledgments: Work at Brookhaven National Laboratory was carried out under Contract No. DE-AC02-98CH10886 and DE-SC0012704 with the U.S. Department of Energy and supported by its Office of Basic Energy Sciences, Division of Chemical Sciences, Geosciences and Biosciences.

Translational-to-vibrational energy transfer (ET) from a hyperthermal H atom to ambient SO₂ was characterized using time-resolved Fourier transform infrared emission spectroscopy. Vibrational excitation of SO₂, following collisions with H atoms containing 59 kcal/mol of kinetic energy, generated from the 193 nm photolysis of HBr, is detected in two distinct energy distributions: one with excitation predominantly at the fundamental vibrational levels is attributable to classical impulsive collisions, while the other, accounting for 80% of the excited SO₂ with vibrational energy as high as 14,000 cm⁻¹, is proposed to arise from the formation of a transient reactive-complex during the collision. The cross-section for this super ET collision is determined to be 0.53±0.05 Ų, or roughly 2% of all hard sphere collisions. This observation reveals that in collisions between a hyperthermal atom and an ambient molecule, for which a reactive-complex exists on the potential energy surface, a large quantity of translational energy can be transferred to the molecule with high efficiency.

Second-order vibrational perturbation theory (VPT2) is an enormously successful and well-established theory for treating anharmonic effects on the vibrational levels of semi-rigid molecules. Partially as a consequence of the fact that the theory is exact for the Morse potential (which provides an appropriate qualitative model for stretching anharmonicity), VPT2 calculations for such systems with appropriate ab initio potential functions tend to give fundamental and overtone levels that fall within a handful of wavenumbers of experimentally measured positions. As a consequence, the next non-vanishing level of perturbation theory - VPT4 - offers only slight improvements over VPT2 and is not practical for most calculations since it requires information about force constants up through sextic. However, VPT4 (as well as VPT2) can be used for other applications such as the next vibrational correction to rotational constants (the "gammas") and other spectroscopic parameters. In addition, the marriage of VPT with the semi-classical transition state theory of Miller (SCTST) has recently proven to be a powerful and accurate treatment for chemical kinetics. In this talk, VPT4-based SCTST tunneling probabilities and cumulative reaction probabilities are give for the first time for selected low-dimensional model systems. The prospects for VPT4, both practical and intrinsic, will also be discussed.