The retinal protonated Schiff-base (RPSB) in its all-trans form is found in bacterial rhodopsins, whereas visual rhodopsin proteins host 11-cis RPSB. In both cases, photoexcitation initiates fast isomerization of the retinal chromophore, leading to proton transport, storage of chemical energy or signaling. It is an unsolved problem, to which degree this is due to protein interactions or intrinsic RPSB quantum properties. Here, we report on time-resolved action-spectroscopy studies, which show, that upon photoexcitation, cis isomers of RPSB have an almost barrierless fast 400 fs decay, whereas all-trans isomers exhibit a barrier-controlled slow 3 ps decay. Moreover, formation of the 11-cis isomer is greatly favored for all-trans RPSB when isolated. The very fast photoresponse of visual photoreceptors is thus directly related to intrinsic retinal properties, whereas bacterial rhodopsins tune the excited state potential-energy surface to lower the barrier for particular double-bond isomerization, thus changing both the timescale and specificity of the photoisomerization.

While action spectroscopy of cold molecular ions is a well-established technique to provide vibrationally resolved absorption features, fluorescence experiments are still challenging. Here we report the fluorescence spectra of pyronin-Y and resorufin ions at 100 K using a newly constructed setup. Spectra narrow upon cooling, and the emission maxima blueshift. Temperature effects are attributed to the population of vibrational excited levels in S 1 , and that frequencies are lower in S 1 than in S 0 . This picture is supported by calculated spectra based on a Franck−Condon model that not only predicts the observed change in maximum, but also assigns Franck−Condon active vibrations. In-plane vibrational modes that preserve the mirror plane present in both S 0 and S 1 of resorufin and pyronin Y account for most of the observed vibrational bands. Finally, at low temperatures, it is important to pick an excitation wavelength as far to the red as possible to not reheat the ions.

Here, we present a new instrument named LUNA2 (LUminescence iNstrument in Aarhus 2), which is purpose-built to measure dispersed fluorescence spectra of gaseous ions produced by electrospray ionization and cooled to low temperatures (<100 K). LUNA2 is, as an earlier room-temperature setup (LUNA), optimized for a high collection efficiency of photons and includes improvements based on our operational experience with LUNA. The fluorescence cell is a cylindrical Paul trap made of copper with a hole in the ring electrode to permit laser light to interact with the trapped ions, and one end-cap electrode is a mesh grid combined with an aspheric condenser lens. The entrance and exit electrodes are both in physical contact with the liquid-nitrogen cooling unit to reduce cooling times. Mass selection is done in a two-step scheme where, first, high-mass ions are ejected followed by low-mass ions according to the Mathieu stability region. This scheme may provide a higher mass resolution than when only one DC voltage is used. Ions are irradiated by visible light delivered from a nanosecond 20-Hz pulsed laser, and dispersed fluorescence is measured with a spectrometer combined with an iCCD camera that allows intensification of the signal for a short time interval. LUNA2 contains an additional Paul trap that can be used for mass selection before ions enter the fluorescence cell, which potentially is relevant to diminishing RF heating in the cold trap. Successful operation of the setup is demonstrated from experiments with rhodamine dyes and oxazine-4, and spectral changes with temperature are identified.

A vibrating stretched string is one of the most fundamental physical systems where non-linear effects play a central role. We describe results obtained with a new, dedicated experimental setup that allows unverified details of the dynamics of this classical problem to be addressed experimentally. The setup uniquely allows simultaneous registration of the transverse motion of the string as well as the longitudinal tension in the string during oscillation over extended times. The explicit non-linear connection between the string tension and the transverse oscillation amplitude is thus directly experimentally determined, allowing us to distinguish between previously reported diverging descriptions. For forced vibrations, frequency response curves for the transverse amplitudes are obtained, and found to be in good agreement with predictions from a simplified model, while the model only qualitatively reproduces the observed phases of transverse oscillation relative to the phase of the driving force. A frequency analysis of the observed string tension reveals a richer dynamics than anticipated from a simple model, and from the observed frequency response curves alone. For free oscillations, the setup is used to demonstrate the variation of the normal mode frequency with the transverse amplitudes. The reported setup is well-suited as a demonstration experiment on non-linear oscillatory phenomena, as well as a student project in either introductory or advanced mechanics teaching.

Here we uncover the direct effect of a high electric field on the absorption by the Green Fluorescent Protein chromophore anion isolated in vacuo based on gas-phase action spectroscopy. Betaine is a strong molecular dipole that creates an electric field of ∼70 MV/cm when attached to the ion at the phenolate oxygen, more than half the actual field from the protein matrix and pointing in the same direction. Nevertheless, the shift in absorption is limited (0.08 eV), supporting earlier conclusions, but subject to much debate, that the protein is rather innocent in perturbing the transition energy. The betaine complexes are readily made by electrospray ionization and in contrast to the bare ions, they dissociate after one-photon absorption. Also, electron detachment is not an open channel complicating the bare ion case. As steric constraints are absent in vacuo, the possibility of turning on fluorescence by an electric field can be tested from experiments on complexes with betaine.

We have investigated the photo-stability of pristine and super-hydrogenated pyrene cations (C 16 H + 10+m , m = 0, 6, or 16) by means of gas-phase action spectroscopy. Optical absorption spectra and photo-induced dissociation mass spectra are presented. By measuring the yield of mass-selected photo-fragment ions as a function of laser pulse intensity, the number of photons (and hence the energy) needed for fragmentation of the carbon backbone was determined. Backbone fragmentation of pristine pyrene ions (C 16 H + 10 ) requires absorption of three photons of energy just below 3 eV, whereas super-hydrogenated hexahydropyrene (C 16 H + 16 ) must absorb two such photons and fully hydrogenated hexadecahydropyrene (C 16 H + 26 ) only a single photon. These results are consistent with previously reported dissociation energies for these ions. Our experiments clearly demonstrate that the increased heat capacity from the additional hydrogen atoms does not compensate for the weakening of the carbon backbone when pyrene is hydrogenated. In photodissociation regions, super-hydrogenated Polycyclic Aromatic Hydrocarbons (PAHs) have been proposed to serve as catalysts for H 2 -formation. Our results indicate that carbon backbone fragmentation may be a serious competitor to H 2 -formation at least for small hydrogenated PAHs like pyrene.

The UV-visible absorption of retinal in its protonated Schiff-base form is studied in the gas phase. In particular, transitions to highly-excited electronic states, S, in the all-trans and 11-cis forms are considered, and several new states are discovered. Their positions and strengths are compared to state of the art quantum calculations. The location of these states are particularly important when new fs pump-probe experiments are designed to investigate the fast excited-state dynamics of retinal chromophores.

We report the absorption profile of isolated Flavin Adenine Dinucleotide (FAD) mono-anions recorded using photo-induced dissociation action spectroscopy. In this charge state, one of the phosphoric acid groups is deprotonated and the chromophore itself is in its neutral oxidized state. These measurements cover the first four optical transitions of FAD with excitation energies from 2.3 to 6.0 eV (210-550 nm). The S → S transition is strongly blue shifted relative to aqueous solution, supporting the view that this transition has a significant charge-transfer character. The remaining bands are close to their solution-phase positions. This confirms that the large discrepancy between quantum chemical calculations of vertical transition energies and solution-phase band maxima cannot be explained by solvent effects. We also report the luminescence spectrum of FAD mono-anions in vacuo. The gas-phase Stokes shift for S is 3000 cm, which is considerably larger than any previously reported for other molecular ions and consistent with a significant displacement of the ground and excited state potential energy surfaces. Consideration of the vibronic structure is thus essential for simulating the absorption and luminescence spectra of flavins.