Serial X-ray crystallography at free-electron lasers allows to solve biomolecular structures from sub-micron-sized crystals. However, beam time at these facilities is scarce, and involved sample delivery techniques are required. On the other hand, rotation electron diffraction (MicroED) has shown great potential as an alternative means for protein nanocrystallography. Here, we present a method for serial electron diffraction of protein nanocrystals combining the benefits of both approaches. In a scanning transmission electron microscope, crystals randomly dispersed on a sample grid are automatically mapped, and a diffraction pattern at fixed orientation is recorded from each at a high acquisition rate. Dose fractionation ensures minimal radiation damage effects. We demonstrate the method by solving the structure of granulovirus occlusion bodies and lysozyme to resolutions of 1.55 Å and 1.80 Å, respectively. Our method promises to provide rapid structure determination for many classes of materials with minimal sample consumption, using readily available instrumentation.

The dynamics of the photoinduced commensurate to incommensurate charge density wave (CDW) phase transition in 4H b -TaSe2 are investigated by femtosecond electron diffraction. In the perturbative regime the CDW reforms on a 150 ps timescale, which is two orders of magnitude slower than in other transition-metal dichalcogenides. We attribute this to a weak coupling between the CDW carrying T layers and thus demonstrate the importance of three-dimensionality for the existence of CDWs. With increasing optical excitation the phase transition is achieved showing a second order character in contrast to the first order behavior in thermal equilibrium.Reduced dimensionality seems to be a decisive property governing phenomena like high temperature superconductivity and charge density wave (CDW) formation. The latter is typical for quasi one-or two-dimensional metals where, in the ground state, the crystal displays a static periodic modulation of the conduction electron density accompanied by a periodic lattice displacement (PLD), both characterized by the wave vector q CDW [1]. The standard theory that describes the appearance of this macroscopic quantum state was formulated by Peierls [2]. By considering a one dimensional metal he has shown that the divergent static electronic susceptibility at a wave vector q = 2k F gives rise to an instability of the electronic system against perturbations at this wave vector. This so called Fermi surface nesting lowers the frequencies of q = 2k F phonons, which will eventually evolve into a static lattice displacement. From that band gaps at ± k F result, which reduce the total electronic energy. If the elastic energy cost to modulate atomic positions is lower than the electronic energy gain, the CDW state is the preferred ground state. Recently, this classical picture has been challenged [3][4][5], since (a) the nesting condition derived from the topology of the Fermi surface (2k F ) and the observed CDW modulation vectors (q CDW ) are not generally equal, and (b) the diverging susceptibility at q = 2k F is exceedingly fragile with respect to temperature, scattering or imperfect nesting [3]. Contrasting the standard Fermi surface nesting scenario the transition from the metallic into a CDW state was argued to occur due to strong qdependent electron-phonon coupling [3], particularly in transition-metal dichalcogenides [4].Only the concerted interplay of electronic and lattice degrees of freedom make the CDW formation possible. The two modulations can be individually examined by scanning tunneling microscopy [6], angular resolved photoemission spectroscopy [5] and electron, x-ray or neutron diffraction techniques [7]. In thermal equilibrium, the gap in the electronic spectrum and the atomic displacement amplitude (A) present different projections of the same order parameter [1]. Adding femtosecond temporal resolution to the experiment enables investigation of their dynamical behavior. Since the electronic system can be perturbed on timescales much faster than the characteristic lattic...

We demonstrate a method of time-stamping Radio Frequency compressed electron bunches for Ultrafast Electron Diffraction experiments in the sub-pC regime. We use an in-situ ultra-stable photo-triggered streak camera to directly track the time of arrival of each electron pulse and correct for the timing jitter in the radio frequency synchronization. We show that we can correct for timing jitter down to 30 fs root-mean-square with minimal distortion to the diffraction patterns, and performed a proof-of-principle experiment by measuring the ultrafast electron-phonon coupling dynamics of silicon

We have developed a compact streak camera suitable for measuring the duration of highly charged subrelativistic femtosecond electron bunches with an energy bandwidth in the order of 0.1%, as frequently used in ultrafast electron diffraction ͑UED͒ experiments for the investigation of ultrafast structural dynamics. The device operates in accumulation mode with 50 fs shot-to-shot timing jitter, and at a 30 keV electron energy, the full width at half maximum temporal resolution is 150 fs. Measured durations of pulses from our UED gun agree well with the predictions from the detailed charged particle trajectory simulations.

Electron ptychography has seen a recent surge of interest for phase sensitive imaging at atomic or near-atomic resolution. However, applications are so far mainly limited to radiation-hard samples, because the required doses are too high for imaging biological samples at high resolution. We propose the use of non-convex Bayesian optimization to overcome this problem, and show via numerical simulations that the dose required for successful reconstruction can be reduced by two orders of magnitude compared to previous experiments. As an important application we suggest to use this method for imaging single biological macromolecules at cryogenic temperatures and demonstrate 2D single-particle reconstructions from simulated data with a resolution up to 5.4 Å at a dose of 20e −/Å2. When averaging over only 30 low-dose datasets, a 2D resolution around 3.5 Å is possible for macromolecular complexes even below 100 kDa. With its independence from the microscope transfer function, direct recovery of phase contrast, and better scaling of signal-to-noise ratio, low-dose cryo electron ptychography may become a promising alternative to Zernike phase-contrast microscopy.

Divalent metal cations can play a role in protein aggregation diseases, including cataract. Here we compare the aggregation of human γS-crystallin, a key structural protein of the eye lens, via mutagenesis, ultraviolet light damage, and the addition of metal ions. All three aggregation pathways result in globular, amorphous-looking structures that do not elongate into fibers. We also investigate the molecular mechanism underlying copper(II)induced aggregation. This work was motivated by the observation that zinc(II)-induced aggregation of γS-crystallin is driven by intermolecular bridging of solvent-accessible cysteine residues, while in contrast, copper(II)-induced aggregation of this protein is exacerbated by the removal of solvent-accessible cysteines via mutation. Here we find that copper(II)-induced aggregation results from a complex mechanism involving multiple interactions with the protein. The initial protein−metal interactions result in the reduction of Cu(II) to Cu(I) with concomitant oxidation of γScrystallin. In addition to the intermolecular disulfides that represent a starting point for aggregation, intramolecular disulfides also occur in the cysteine loop, a region of the N-terminal domain that was previously found to mediate the early stages of cataract formation. This previously unobserved ability of γS-crystallin to transfer disulfides intramolecularly suggests that it may serve as an oxidation sink for the lens after glutathione levels have become depleted during aging. γS-Crystallin thus serves as the last line of defense against oxidation in the eye lens, a result that underscores the chemical functionality of this protein, which is generally considered to play a purely structural role.

Through combined three-dimensional electromagnetic and particle tracking simulations we demonstrate a THz driven electron streak camera featuring a temporal resolution on the order of a femtosecond. The ultrafast streaking field is generated in a resonant THz sub-wavelength antenna which is illuminated by an intense single-cycle THz pulse. Since electron bunches and THz pulses are generated with parts of the same laser system, synchronization between the two is inherently guaranteed.

Base-pairing stability in DNA-gold nanoparticle (DNA-AuNP) multimers along with their dynamics under different electron beam intensities was investigated with in-liquid transmission electron microscopy (in-liquid TEM). Multimer formation was triggered by hybridization of DNA oligonucleotides to another DNA strand (Hyb-DNA) related to the concept of DNA origami. We analyzed the degree of multimer formation for a number of samples and a series of control samples to determine the specificity of the multimerization during the TEM imaging. DNA-AuNPs with Hyb-DNA showed an interactive motion and assembly into 1D structures once the electron beam intensity exceeds a threshold value. This behavior was in contrast with control studies with noncomplementary DNA linkers where statistically significantly reduced multimerization was observed and for suspensions of citrate-stabilized AuNPs without DNA, where we did not observe any significant motion or aggregation. These findings indicate that DNA base-pairing interactions are the driving force for multimerization and suggest a high stability of the DNA base pairing even under electron exposure.