Determinants of synapse diversity revealed by super-resolution quantal transmission and active zone imaging

Neural circuit function depends on the pattern of synaptic connections between neurons and the strength of those connections. Synaptic strength is determined by both postsynaptic sensitivity to neurotransmitter and the presynaptic probability of action potential evoked transmitter release (Pr). Whereas morphology and neurotransmitter receptor number indicate postsynaptic sensitivity, presynaptic indicators and the mechanism that sets Pr remain to be defined. To address this, we developed QuaSOR, a super-resolution method for determining Pr from quantal synaptic transmission imaging at hundreds of glutamatergic synapses at a time. We mapped the Pr onto super-resolution 3D molecular reconstructions of the presynaptic active zones (AZs) of the same synapses at the Drosophila larval neuromuscular junction (NMJ). We find that Pr varies greatly between synapses made by a single axon, quantify the contribution of key AZ proteins to Pr diversity and find that one of these, Complexin, suppresses spontaneous and evoked transmission differentially, thereby generating a spatial and quantitative mismatch between release modes. Transmission is thus regulated by the balance and nanoscale distribution of release-enhancing and suppressing presynaptic proteins to generate high signal-to-noise evoked transmission.

Thermal Drift Correction for Laboratory Nano Computed Tomography via Outlier Elimination and Feature Point Adjustment

Thermal drift of nano-computed tomography (CT) adversely affects the accurate reconstruction of objects. However, feature-based reference scan correction methods are sometimes unstable for images with similar texture and low contrast. In this study, based on the geometric position of features and the structural similarity (SSIM) of projections, a rough-to-refined rigid alignment method is proposed to align the projection. Using the proposed method, the thermal drift artifacts in reconstructed slices are reduced. Firstly, the initial features are obtained by speeded up robust features (SURF). Then, the outliers are roughly eliminated by the geometric position of global features. The features are refined by the SSIM between the main and reference projections. Subsequently, the SSIM between the neighborhood images of features are used to relocate the features. Finally, the new features are used to align the projections. The two-dimensional (2D) transmission imaging experiments reveal that the proposed method provides more accurate and robust results than the random sample consensus (RANSAC) and locality preserving matching (LPM) methods. For three-dimensional (3D) imaging correction, the proposed method is compared with the commonly used enhanced correlation coefficient (ECC) method and single-step discrete Fourier transform (DFT) algorithm. The results reveal that proposed method can retain the details more faithfully.

Comparative Study of Modified Mueller Matrix Transformation and Polar Decomposition Parameters for Transmission and Backscattering Tissue Polarimetries

Mueller matrix polarimetry is widely used in biomedical studies and applications, for it can provide abundant microstructural information about tissues. Recently, several methods have been proposed to decompose the Mueller matrix into groups of parameters related to specific optical properties which can be used to reveal the microstructural information of tissue samples more clearly and quantitatively. In this study, we thoroughly compare the differences among the parameters derived from the Mueller matrix polar decomposition (MMPD) and Mueller matrix transformation (MMT), which are two popular methods in tissue polarimetry studies and applications, while applying them on different tissue samples for both backscattering and transmission imaging. Based on the Mueller matrix data obtained using the setups, we carry out a comparative analysis of the parameters derived from both methods representing the same polarization properties, namely depolarization, linear retardance, fast axis orientation and diattenuation. IN particular, we propose several modified MMT parameters, whose abilities are also analyzed for revealing the information about the specific type of tissue samples. The results presented in this study evaluate the applicability of the original and modified MMT parameters, then give the suggestions for appropriate parameter selection in tissue polarimetry, which can be helpful for future biomedical and clinical applications.

DMD-based hyperspectral microscopy with flexible multiline parallel scanning

As one of the most common hyperspectral microscopy (HSM) techniques, line-scanning HSM is currently utilized in many fields. However, its scanning efficiency is still considered to be inadequate since many biological and chemical processes occur too rapidly to be captured. Accordingly, in this work, a digital micromirror device (DMD) based on microelectromechanical systems (MEMS) is utilized to demonstrate a flexible multiline scanning HSM system. To the best of our knowledge, this is the first line-scanning HSM system in which the number of scanning lines N can be tuned by simply changing the DMD’s parallel scanning units according to diverse applications. This brilliant strategy of effortless adjustability relies only on on-chip scanning methods and totally exploits the benefits of parallelization, aiming to achieve nearly an N-time improvement in the detection efficiency and an N-time decrease in the scanning time and data volume compared with the single-line method under the same operating conditions. To validate this, we selected a few samples of different spectral wavebands to perform reflection imaging, transmission imaging, and fluorescence imaging with varying numbers of scanning lines. The results show the great potential of our DMD-based HSM system for the rapid development of cellular biology, material analysis, and so on. In addition, its on-chip scanning process eliminates the inherent microscopic architecture, making the whole system compact, lightweight, portable, and not subject to site constraints.

Analysis and Mapping of Detailed Inner Information of Crystalline Grain by Wavelength-Resolved Neutron Transmission Imaging with Individual Bragg-Dip Profile-Fitting Analysis

As a new method for evaluating single crystals and oligocrystals, pulsed neutron Bragg-dip transmission analysis/imaging method is being developed. In this study, a single Bragg-dip profile-fitting analysis method was newly developed, and applied for analyzing detailed inner information in a crystalline grain position-dependently. In the method, the spectrum profile of a single Bragg-dip is analyzed at each position over a grain. As a result, it is expected that changes in crystal orientation, mosaic spread angle and thickness of a perfect crystal can be evaluated from the wavelength, the width and the integrated intensity of the Bragg-dip, respectively. For confirming this effectiveness, the method was applied to experimental data of position-dependent Bragg-dip transmission spectra of a Si-steel plate consisting of oligocrystals. As a result, inner information of multiple crystalline grains could be visualized and evaluated. The small change in crystal orientation in a grain, about 0.4°, could be observed by imaging the Bragg-dip wavelengths. By imaging the Bragg-dip widths, both another grain and mosaic block in a grain were detected. Furthermore, imaging results of the integrated intensities of Bragg-dips were consistent with the results of Bragg-dip width imaging. These small crystallographic changes have not been observed and visualized by previous Bragg-dip analysis methods.

X-ray fan beam coded aperture transmission and diffraction imaging for fast material analysis

X-ray transmission imaging has been used in a variety of applications for high-resolution measurements based on shape and density. Similarly, X-ray diffraction (XRD) imaging has been used widely for molecular structure-based identification of materials. Combining these X-ray methods has the potential to provide high-resolution material identification, exceeding the capabilities of either modality alone. However, XRD imaging methods have been limited in application by their long measurement times and poor spatial resolution, which has generally precluded combined, rapid measurements of X-ray transmission and diffraction. In this work, we present a novel X-ray fan beam coded aperture transmission and diffraction imaging system, developed using commercially available components, for rapid and accurate non-destructive imaging of industrial and biomedical specimens. The imaging system uses a 160 kV Bremsstrahlung X-ray source while achieving a spatial resolution of ≈ 1 × 1 mm2 and a spectral accuracy of > 95% with only 15 s exposures per 150 mm fan beam slice. Applications of this technology are reported in geological imaging, pharmaceutical inspection, and medical diagnosis. The performance of the imaging system indicates improved material differentiation relative to transmission imaging alone at scan times suitable for a variety of industrial and biomedical applications.

Potential of Dual and Multi Energy XRT and CT Analyses on Iron Formations

Dual and multi energy X-ray transmission imaging (DE-/ME-XRT) are powerful tools to acquire quantitative material characteristics of diverse samples without destruction. As those X-ray imaging techniques are based on the projection onto the imaging plane, only two-dimensional data can be obtained. To acquire three-dimensional information and a complete examination on topology and spatial trends of materials, computed tomography (CT) can be used. In combination, these methods may offer a robust non-destructive testing technique for research and industrial applications. For example, the iron ore mining and processing industry requires the ratio of economic iron minerals to siliceous waste material for resource and reserve estimations, and for efficient sorting prior to beneficiation, to avoid equipment destruction due to highly abrasive quartz. While XRT provides information concerning the thickness, areal density and mass fraction of iron and the respective background material, CT may deliver size, distribution and orientation of internal structures. Our study shows that the data provided by XRT and CT is reliable and, together with data processing, can be successfully applied for distinguishing iron oxide rich parts from waste. Furthermore, heavy element bearing minerals such as baryte, uraninite, galena and monazite can be detected.