Unravelling the interaction of biological macromolecules with ligands and substrates at high spatial and temporal resolution remains a major challenge in structural biology. The development of serial crystallography methods at X-ray free-electron lasers and subsequently at synchrotron light sources allows new approaches to tackle this challenge. Here, a new polyimide tape drive designed for mix-and-diffuse serial crystallography experiments is reported. The structure of lysozyme bound by the competitive inhibitor chitotriose was determined using this device in combination with microfluidic mixers. The electron densities obtained from mixing times of 2 and 50 s show clear binding of chitotriose to the enzyme at a high level of detail. The success of this approach shows the potential for high-throughput drug screening and even structural enzymology on short timescales at bright synchrotron light sources.
Here we present a new approach to diffraction imaging of amyloid fibrils, combining a free-standing graphene support and single nanofocused X-ray pulses of femtosecond duration from an X-ray free-electron laser. Due to the very low background scattering from the graphene support and mutual alignment of filaments, diffraction from tobacco mosaic virus (TMV) filaments and amyloid protofibrils is obtained to 2.7 Å and 2.4 Å resolution in single diffraction patterns, respectively. Some TMV diffraction patterns exhibit asymmetry that indicates the presence of a limited number of axial rotations in the XFEL focus. Signal-to-noise levels from individual diffraction patterns are enhanced using computational alignment and merging, giving patterns that are superior to those obtainable from synchrotron radiation sources. We anticipate that our approach will be a starting point for further investigations into unsolved structures of filaments and other weakly scattering objects.
9 December 2019 in Journal of Lightwave Technology
We present the fabrication and characterization of optical fiber reflection probes based on surface-enhanced Raman scattering (SERS) and micro-lensed multimode fibers. For the SERS substrate, a nano-sphere lithography method is used. Comparison of SERS measurements with three different fiber probe configurations is presented. The proposed optimized structure shows a five times increase in SERS signal for dried Rhodamine 6G (R6G) and two times for aqueous R6G in comparison to a standard reflection configuration. Measurements of different concentrations of R6G in a water solution with an limit of detection (LOD) equal to 10 −7 M are demonstrated.
We present the fabrication and characterization of a novel sensing configuration based on surface-enhanced Raman scattering (SERS) and two micro-lensed optical fibers. The first micro-lensed fiber is used to excite surface plasmon resonance in a gold film deposited over a mono-layer of nano-sphere surface (AuFON), and the second lensed fiber is used to collect the SERS signal. The sensing capabilities of the fabricated device are demonstrated by measuring different concentrations of Rhodamine 6G in a water solution.
Visible and near-infrared spectroscopy are widely used for sensing applications but suffer from poor signal-to-noise ratios for the detection of compounds with low concentrations. Enhancement by surface plasmon resonance is a popular technique that can be utilized to increase the signal of absorption spectroscopy due to the increased near-field created close to the plasmons. Despite interest in surface-enhanced infrared absorption spectroscopy (SEIRAS), the method is usually applied in lab setups rather than real-life sensing situations. This study aimed to achieve enhanced absorption from plasmons on a fiber-optic probe and thus move closer to applications of SEIRAS. A tapered coreless fiber coated with a 100 nm Au film supported signal enhancement at visible wavelengths. An increase in absorption was shown for two dyes spanning concentrations from 5 × 10−8 mol/L to 8 × 10−4 mol/L: Rhodamine 6G and Crystal Violet. In the presence of the Au film, the absorbance signal was 2–3 times higher than from an identically tapered uncoated fiber. The results confirm that the concept of SEIRAS can be implemented on an optical fiber probe, enabling enhanced signal detection in remote sensing applications.