Mutations to R. sphaeroides Reaction Center Perturb Energy Levels and Vibronic Coupling but Not Observed Energy Transfer Rates.

The bacterial reaction center is capable of both efficiently collecting and quickly transferring energy within the complex; therefore, the reaction center serves as a convenient model for both energy transfer and charge separation. To spectroscopically probe the interactions between the electronic excited states on the chromophores and their intricate relationship with vibrational motions in their environment, we examine coherences between the excited states. Here, we investigate this question by introducing a series of point mutations within 12 Å of the special pair of bacteriochlorophylls in the Rhodobacter sphaeroides reaction center. Using two-dimensional spectroscopy, we find that the time scales of energy transfer dynamics remain unperturbed by these mutations. However, within these spectra, we detect changes in the mixed vibrational-electronic coherences in these reaction centers. Our results indicate that resonance between bacteriochlorophyll vibrational modes and excitonic energy gaps promote electronic coherences and support current vibronic models of photosynthetic energy transfer.

Dynamic localization of electronic excitation in photosynthetic complexes revealed with chiral two-dimensional spectroscopy.

Time-resolved ultrafast optical probes of chiral dynamics provide a new window allowing us to explore how interactions with such structured environments drive electronic dynamics. Incorporating optical activity into time-resolved spectroscopies has proven challenging because of the small signal and large achiral background. Here we demonstrate that two-dimensional electronic spectroscopy can be adapted to detect chiral signals and that these signals reveal how excitations delocalize and contract following excitation. We dynamically probe the evolution of chiral electronic structure in the light-harvesting complex 2 of purple bacteria following photoexcitation by creating a chiral two-dimensional mapping. The dynamics of the chiral two-dimensional signal directly reports on changes in the degree of delocalization of the excitonic states following photoexcitation. The mechanism of energy transfer in this system may enhance transfer probability because of the coherent coupling among chromophores while suppressing fluorescence that arises from populating delocalized states. This generally applicable spectroscopy will provide an incisive tool to probe ultrafast transient molecular fluctuations that are obscured in non-chiral experiments.

Probing energy transfer events in the light harvesting complex 2 (LH2) of Rhodobacter sphaeroides with two-dimensional spectroscopy.

Excitation energy transfer events in the photosynthetic light harvesting complex 2 (LH2) of Rhodobacter sphaeroides are investigated with polarization controlled two-dimensional electronic spectroscopy. A spectrally broadened pulse allows simultaneous measurement of the energy transfer within and between the two absorption bands at 800 nm and 850 nm. The phased all-parallel polarization two-dimensional spectra resolve the initial events of energy transfer by separating the intra-band and inter-band relaxation processes across the two-dimensional map. The internal dynamics of the 800 nm region of the spectra are resolved as a cross peak that grows in on an ultrafast time scale, reflecting energy transfer between higher lying excitations of the B850 chromophores into the B800 states. We utilize a polarization sequence designed to highlight the initial excited state dynamics which uncovers an ultrafast transfer component between the two bands that was not observed in the all-parallel polarization data. We attribute the ultrafast transfer component to energy transfer from higher energy exciton states to lower energy states of the strongly coupled B850 chromophores. Connecting the spectroscopic signature to the molecular structure, we reveal multiple relaxation pathways including a cyclic transfer of energy between the two rings of the complex.

Timescales of Coherent Dynamics in the Light Harvesting Complex 2 (LH2) of Rhodobacter sphaeroides.

The initial dynamics of energy transfer in the light harvesting complex 2 from Rhodobacter sphaeroides were investigated with polarization controlled two-dimensional spectroscopy. This method allows only the coherent electronic motions to be observed revealing the timescale of dephasing among the excited states. We observe persistent coherence among all states and assign ensemble dephasing rates for the various coherences. A simple model is utilized to connect the spectroscopic transitions to the molecular structure, allowing us to distinguish coherences between the two rings of chromophores and coherences within the rings. We also compare dephasing rates between excited states to dephasing rates between the ground and excited states, revealing that the coherences between excited states dephase on a slower timescale than coherences between the ground and excited states.

Energy Transfer Observed in Live Cells Using Two-Dimensional Electronic Spectroscopy.

Two-dimensional electronic spectroscopy (2DES) elucidates electronic structure and dynamics on a femtosecond time scale and has proven to be an incisive tool for probing congested linear spectra of biological systems. However, samples that scatter light intensely frustrate 2DES analysis, necessitating the use of isolated protein chromophore complexes when studying photosynthetic energy transfer processes. We present a method for conducting 2DES experiments that takes only seconds to acquire thousands of 2DES spectra and permits analysis of highly scattering samples, specifically whole cells of the purple bacterium Rhodobacter sphaeroides. These in vivo 2DES experiments reveal similar timescales for energy transfer within the antennae complex (light harvesting complex 2, LH2) both in the native photosynthetic membrane environment and in isolated detergent micelles.

Two-dimensional spectroscopy can distinguish between decoherence and dephasing of zero-quantum coherences.

Recent experiments on a variety of photosynthetic antenna systems have revealed that coherences among electronic states persist longer than previously anticipated. In an ensemble measurement, the observed dephasing of a coherent state can occur because of either disorder across the ensemble or decoherence from interactions with the bath. Distinguishing how much such disorder affects the experimentally observed dephasing rate is paramount for understanding the role that quantum coherence may play in energy transfer through these complexes. Here, we show that two-dimensional electronic spectra can distinguish between the limiting cases of homogeneous dephasing (decoherence) and inhomogeneous dephasing by examining how the quantum beat frequency changes within a cross peak. For the antenna complex LH2 isolated from Rhodobacter sphaeroides , we find that dephasing of the coherence between the B850 and B800 rings arises predominantly from inhomogeneity. In contrast, within the Fenna-Matthews-Olson (FMO) complex from Chlorobium tepidum , dephasing of the coherence between the first two excitons appears quite homogeneous. Thus, the observed dephasing rate sets an upper bound on decoherence for the LH2 complex while establishing both an upper and lower bound for the FMO complex.

Continuing professional development: the missing link.

Attending formal course-based learning forms a major part of continuing professional development for physiotherapists. There is a vast array of formal courses available to physiotherapists each holding the tantalising prospect of improving knowledrge, skills and patient outcomes. However, educational evidence suggests that, while participation in traditionally organised workshops and conferences improves knowledge and practice behaviours of the individual attendee, there is no corollary improvement in patient outcomes. This paper discusses reasons why formal course-based learning has yet to be successful at improving the patient outcomes of those who participate. Suggestions and strategies for reconceptualising this aspect of continuing professional development are provided.

Single-shot ultrabroadband two-dimensional electronic spectroscopy of the light-harvesting complex LH2.

Here we present two-dimensional (2D) electronic spectra of the light-harvesting complex LH2 from purple bacteria using coherent pulses with bandwidth of over 100 nm FWHM. This broadband excitation and detection has allowed the simultaneous capture of both the B800 and B850 bands using a single light source. We demonstrate that one laser pulse is sufficient to capture the entire 2D electronic spectrum with a high signal-to-noise ratio. At a waiting time of 800 fs, we observe population transfer from the B800 to B850 band as manifested by a prominent cross peak. These results will enable observation of the dynamics of biological systems across both ultrafast (<1 ps) and slower (>1 ms) timescales simultaneously.