X-ray ptychography imaging at synchrotron facilities like the Advanced Photon Source (APS) involves controlling instrument hardwares to collect a set of diffraction patterns from overlapping coherent illumination spots on extended samples, managing data storage, reconstructing ptychographic images from acquired diffraction patterns, and providing the visualization of results and feedback. In addition to the complicated workflow, ptychography instrument could produce up to several TB’s of data per second that is needed to be processed in real time. This brings up the need to develop a high performance, robust and user friendly processing software package for ptychographic data analysis. In this paper we present a software framework which provides functionality of visualization, work flow control, and data reconstruction. To accelerate the computation and large datasets process, the data reconstruction part is implemented with three algorithms, ePIE,1 DM2 and LSQML3 using CUDA-C on GPU.
Ptychography has become a popular technique for high-throughput and high-resolution characterization of 2D/3D materials. When objects introduce significantly large phase shifts, a multi-slice model needs to be considered to account for long-distance wave propagation within the sample. Although many groups have demonstrated multi-slice ptychography using specimens that are several times larger than the depth of field (DOF), the benefits of applying the multi-slice ptychography algorithm on small objects within the DOF is rarely discussed. Here we address this question and demonstrate that multi-slice ptychography can play an important role in improving reconstruction quality for continuous objects that are smaller than the DOF.
As a scanning version of coherent diffraction imaging (CDI), X-ray ptychography has become a popular and very successful method for high-resolution quantitative imaging of extended specimens. The requirements of mostly coherent illumination and the scanning mechanism limit the throughput of ptychographic imaging. In this paper, we will introduce the methods we use at the Advanced Photon Source (APS) to achieve highthroughput ptychography by optimizing the parameters of the illumination beam. One work we have done is increasing the illumination flux by using a double-multilayer monochromator (DMM) optics with about 0.8% bandwidth. Compared with our double-crystal monochromator (DCM) optics with 0.01% bandwidth, this DMM optics provides around 20 times more flux. A multi-wavelength reconstruction method has been implemented to deal with the consequential degraded temporal coherence from such an illumination to ensure high-quality reconstruction. In the other work, we adopt a novel use of at-top focusing optics to generate a at-top beam with the diameter of about 1.5 μm on the focal plane. The better uniformity of the probe and the large beam size allow one to significantly increase the step size in ptychography scans and thereby the imaging efficiency.
Among different techniques based on x-ray nanoimaging, ptychography has become a popular tool to study specimens at nanometer-scale resolution without the need of using high-resolution optics that requires very stringent manufacturing processes. This high-resolution imaging method is compatible with other imaging modalities acquired in scanning microscopy. At the Advance Photon Source (APS), we have developed two fluorescence microscopes for simultaneous ptychography and fluorescence imaging which together provide a powerful technique to study samples in biology, environmental science, and materials science. Combined with different tilted sample projections, such correlative methods can yield high-resolution 3D structural and chemical images. More recent work has been focused on the development of a fast ptychography instrument called the Velociprobe which is built to take advantage of the over 100 times higher coherent flux provided by the coming APS upgrade source. The Velociprobe uses high-bandwidth accurate interferometry and advanced motion controls with fast continuous scanning schemes which are optimized for large-scale samples and 3D high-resolution imaging. This instrument has been demonstrated to obtain sub-10 nm resolution with different high-photon-efficient scanning schemes using fast data acquisition rate up to 3 kHz (currently limited by detector's full continuous frame rate). A ptychographic imaging rate of 100 _m2/second with a sub-20 nm spatial resolution was shown in this paper.
X-ray ptychography has become a standard technique for imaging materials at <10 nanometer spatial resolution. Recent developments have shown its potential in obtaining quantitative images of the 2D/3D structure of large objects at millimeter and centimeter-scale, which requires not only new instrumentation and experiment design, but high-throughput workflow for data processing. At Argonne’s Advanced Photon Source, we imaged an integrated chip with over 600 × 600 µm^2 field of view at sub-20 nm spatial resolution and achieved 3000 Hz data acquisition rate with advanced motion control. Here, we discuss challenges in achieving large-area reconstruction and explore strategies for streamlining data processing. We demonstrate a novel data acquisition scheme that combines the merits of both step scan and (continuous) fly scan. Inaccurate scan position and large beam variation also degrade image quality and need to be corrected during reconstruction.
Motivated by the Advanced Photon Source Upgrade (APS-U), a new hard X-ray microscope called “Velociprobe” has been recently designed and built for fast ptychographic imaging with high spatial resolution. We are addressing the challenges of high-resolution and fast scanning with novel hardware/stage designs, new positioner control designs, and new data acquisition strategies, including the use of high bandwidth interferometric measurements. The use of granite, air-bearing-supported stages provides the necessary long travel ranges for coarse motion to accommodate real samples and variable energy operation while remaining highly stable during fine scanning. Scanning the low-mass zone plate enables high-speed high-precision motion of the probe over the sample. Our primary goal is to use this instrument to demonstrate sub-10 nm spatial resolution ptychography over a 1-square-micron area in under 10 seconds. We have also designed the instrument to take advantage of the upgraded source when the APS-U is completed. This presentation will describe the unique designs and characteristics of this instrument, and some preliminary data obtained during the instrument commission.
X-ray fluorescence offers unparalleled sensitivity for imaging the nanoscale distribution of trace elements in micrometer thick samples, while x-ray ptychography offers an approach to image weakly fluorescing lighter elements at a resolution beyond that of the x-ray lens used. These methods can be used in combination, and in continuous scan mode for rapid data acquisition when using multiple probe mode reconstruction methods. We discuss here the opportunities and limitations of making use of additional information provided by ptychography to improve x-ray fluorescence images in two ways: by using position-error-correction algorithms to correct for scan distortions in fluorescence scans, and by considering the signal-to-noise limits on previously-demonstrated ptychographic probe deconvolution methods. This highlights the advantages of using a combined approach.
Hard X-ray fluorescence (XRF) microscopy offers unparalleled sensitivity for quantitative analysis of most of the trace elements in biological samples, such as Fe, Cu, and Zn. These trace elements play critical roles in many biological processes. With the advanced nano-focusing optics, nowadays hard X-rays can be focused down to 30 nm or below and can probe trace elements within subcellular compartments. However, XRF imaging does not usually reveal much information on ultrastructure, because the main constituents of biomaterials, i.e. H, C, N, and O, have low fluorescence yield and little absorption contrast at multi-keV X-ray energies. An alternative technique for imaging ultrastructure is ptychography. One can record far-field diffraction patterns from a coherently illuminated sample, and then reconstruct the complex transmission function of the sample. In theory the spatial resolution of ptychography can reach the wavelength limit. In this manuscript, we will describe the implementation of ptychography at the Bionanoprobe (a recently developed hard XRF nanoprobe at the Advanced Photon Source) and demonstrate simultaneous ptychographic and XRF imaging of frozen-hydrated biological whole cells. This method allows locating trace elements within the subcellular structures of biological samples with high spatial resolution. Additionally, both ptychographic and XRF imaging are compatible with tomographic approach for 3D visualization.
Hard X-ray fluorescence microscopy is one of the most sensitive techniques to perform trace elemental analysis of
unsectioned biological samples, such as cells and tissues. As the spatial resolution increases beyond sub-micron
scale, conventional sample preparation method, which involves dehydration, may not be sufficient for preserving
subcellular structures in the context of radiation-induced artifacts. Imaging of frozen-hydrated samples under
cryogenic conditions is the only reliable way to fully preserve the three dimensional structures of the samples while
minimizing the loss of diffusible ions. To allow imaging under this hydrated “natural-state” condition, we have
developed the Bionanoprobe (BNP), a hard X-ray fluorescence nanoprobe with cryogenic capabilities, dedicated to
studying trace elements in frozen-hydrated biological systems. The BNP is installed at an undulator beamline at Life
Sciences Collaboration Access Team at the Advanced Photon Source. It provides a spatial resolution of 30 nm for
fluorescence imaging by using Fresnel zone plates as nanofocusing optics. Differential phase contrast imaging is
carried out in parallel to fluorescence imaging by using a quadrant photodiode mounted downstream of the sample.
By employing a liquid-nitrogen-cooled sample stage and cryo specimen transfer mechanism, the samples are well
maintained below 110 K during both transfer and X-ray imaging. The BNP is capable for automated tomographic
dataset collection, which enables visualization of internal structures and composition of samples in a nondestructive
manner. In this presentation, we will describe the instrument design principles, quantify instrument performance,
and report the early results that were obtained from frozen-hydrated whole cells.
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