In situ/In vivo Optical Microspectroscopy to Probe the Emergence of Morphology.

Morphology governs function. Yet, understanding and controlling the emergence of morphology at the molecular level remains challenging. The difficulty in studying the early stage of morphology formation is due to its stochastic nature both spatially and temporally occurring at the nanoscale. This nature has been particularly detrimental for the application of optical spectroscopy. To overcome this problem, we have been developing new in situ/in vivo optical spectroscopy tools, which are label-free and non-invasive. This account highlights several examples of how optical spectroscopy can become an important tool in studying the birth of morphology.

Unveiling the Configurational Landscape of Carbamate: Paving the Way for Designing Functional Sequence-Defined Polymers.

Carbamate is an emerging class of a polymer backbone for constructing sequence-defined, abiotic polymers. It is expected that new functional materials can be de novo designed by controlling the primary polycarbamate sequence. While amino acids have been actively studied as building blocks for protein folding and peptide self-assembly, carbamates have not been widely investigated from this perspective. Here, we combined infrared (IR), vibrational circular dichroism (VCD), and nuclear magnetic resonance (NMR) spectroscopy with density functional theory (DFT) calculations to understand the conformation of carbamate monomer units in a nonpolar, aprotic environment (chloroform). Compared with amino acid building blocks, carbamates are more rigid, presumably due to the extended delocalization of π-electrons on the backbones. Cis configurations of the amide bond can be energetically stable in carbamates, whereas peptides often assume trans configurations at low energies. This study lays an essential foundation for future developments of carbamate-based sequence-defined polymer material design.

Toward time resolved dynamic light scattering microscopy: Retrieving particle size distributions at high temporal resolutions.

Dynamic light scattering (DLS) is a widely applied technique in multiple scientific and industrial fields for the size characterization of nanoscale objects in solution. While DLS is typically applied to characterize systems under static conditions, the emerging interest in using DLS on temporally evolving systems stimulates the latent need to improve the time resolution of measurements. Herein, we present a DLS microscopy setup (micro-DLS) that can accurately characterize the size of particles from autocorrelation functions built from sub-100 ms time windows, several orders of magnitude faster than previously reported. The system first registers the arrival time of the scattered photons using a time-correlated single photon counting module, which allows the construction of the autocorrelation function for size characterization based on a time window of freely chosen position and width. The setup could characterize both monomodal (60 or 220 nm polystyrene particles; PS) and multimodal size distributions (e.g., mixture of 20 nm LUDOX and 80 nm PS) with high accuracy in a sub-100 ms time window. Notably, the width of the size distribution became narrower as a shorter time window was used. This was attributed to the ability of the system to resolve the sub-ensemble of the broad size distribution, as the broad distribution could be reconstructed by accumulating the distribution obtained by consecutive 80 ms time windows. A DLS system with high temporal resolution will accelerate the expansion of its application toward systems that evolve as a function of time beyond its conventional use on static systems.

Efficient Photoredox Cycles to Control Perylenediimide Self-Assembly.

Photoreduction of perylenediimide (PDI) derivatives has been widely studied for use in photocatalysis, hydrogen evolution, photo-responsive gels, and organic semiconductors. Upon light irradiation, the radical anion (PDI⋅- ) can readily be obtained, whereas further reduction to the dianion (PDI2- ) is rare. Here we show that full 2-electron photoreduction can be achieved using UVC light: 1) in anaerobic conditions by 'direct photoreduction' of PDI aggregates, or 2) by 'indirect photoreduction' in aerobic conditions due to acetone ketyl radicals. The latter strategy is also efficient for other dyes, such as naphthalenediimide (NDI) and methylviologen (MV2+ ). Efficient photoreduction on the minute time-scale using simple LED light in aerobic conditions is attractive for use in dissipative light-driven systems and materials.

Combined in vitro and in vivo investigation of barite microcrystals in Spirogyra (Zygnematophyceae, Charophyta).

We have investigated the biomineralisation of barite ‒a useful proxy for reconstructing paleoproductivity‒ in a freshwater alga, Spirogyra, by combining in vitro and in vivo approaches to unveil the nature of its barite microcrystals. Scanning electron microscope (SEM) and energy-dispersive X-ray spectroscopy (EDXS) observations on simply dried samples revealed that the number and size of barite crystals were related to the barium concentration in the media. Additionally, their morphology showed a crystallographic face (011), which is not normally observed, suggesting the influence of organic molecules on the growth kinetics. The critical point drying method was used to preserve the internal and external structures of Spirogyra cells for SEM imaging. Crystals were found adjacent to the cytoplasmic membrane, near chloroplasts and fibrillary network. In vivo optical microscopy and Raman tweezer microspectroscopy in living cells showed that barite microcrystals are optically visible and follow cytoplasmic streaming. These results led us to propose that barite formation in Spirogyra occurs in the cytoplasm where barium and sulphate are both available: barium supplied non-selectively through the active transport of the divalent cations needed for actin polymerisation, and sulphate because necessary for amino acid biosynthesis in chloroplasts.

Hazardous chemical elements in cleaning cloths, a potential source of microfibres.

Although potentially hazardous chemical elements (e.g., Cu, Cr, Pb, Sb, Ti, Zn) have been studied in clothing textiles, their presence in cleaning textiles is unknown. In this study, 48 cleaning cloth products (consisting of 81 individual samples) purchased in Europe, and consisting of synthetic (petroleum-based), semi-synthetic or natural fibres or combinations of these different types, have been analysed for 16 chemical elements by X-ray fluorescence (XRF) spectrometry. Titanium was detected in most cases (median and maximum concentrations ~3700 and 12,400 mg kg-1, respectively) and Raman microspectroscopy revealed that TiO2 was present as anatase. Barium, Br, Cr, Cu, Fe and Zn were frequently detected over a range of concentrations, reflecting the presence of various additives, and Sb was present at concentrations up to about 200 mg kg-1 in samples containing polyester as catalytic residue from the polymerisation process. Lead was detected as a contaminant in four samples and at concentrations below 10 mg kg-1. Overall, the range of the chemical element profiles and concentrations was similar to those for clothing materials published in the literature, suggesting that broadly the same additives, materials and processes are employed to manufacture cloths and clothing textiles. The mechanisms by which potentially hazardous chemical elements are released into the environment with microfibres or mobilised into soluble or nano-particulate forms remain to be explored.

In situ optical spectroscopy of crystallization: One crystal nucleation at a time.

While crystallization is a ubiquitous and an important process, the microscopic picture of crystal nucleation is yet to be established. Recent studies suggest that the nucleation process can be more complex than the view offered by the classical nucleation theory. Here, we implement single crystal nucleation spectroscopy (SCNS) by combining Raman microspectroscopy and optical trapping induced crystallization to spectroscopically investigate one crystal nucleation at a time. Raman spectral evolution during a single glycine crystal nucleation from water, measured by SCNS and analyzed by a nonsupervised spectral decomposition technique, uncovered the Raman spectrum of prenucleation aggregates and their critical role as an intermediate species in the dynamics. The agreement between the spectral feature of prenucleation aggregates and our simulation suggests that their structural order emerges through the dynamic formation of linear hydrogen-bonded networks. The present work provides a strong impetus for accelerating the investigation of crystal nucleation by optical spectroscopy.