scholarly journals A data reduction and compression description for high throughput time-resolved electron microscopy

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Abhik Datta ◽  
Kian Fong Ng ◽  
Deepan Balakrishnan ◽  
Melissa Ding ◽  
See Wee Chee ◽  
...  

AbstractFast, direct electron detectors have significantly improved the spatio-temporal resolution of electron microscopy movies. Preserving both spatial and temporal resolution in extended observations, however, requires storing prohibitively large amounts of data. Here, we describe an efficient and flexible data reduction and compression scheme (ReCoDe) that retains both spatial and temporal resolution by preserving individual electron events. Running ReCoDe on a workstation we demonstrate on-the-fly reduction and compression of raw data streaming off a detector at 3 GB/s, for hours of uninterrupted data collection. The output was 100-fold smaller than the raw data and saved directly onto network-attached storage drives over a 10 GbE connection. We discuss calibration techniques that support electron detection and counting (e.g., estimate electron backscattering rates, false positive rates, and data compressibility), and novel data analysis methods enabled by ReCoDe (e.g., recalibration of data post acquisition, and accurate estimation of coincidence loss).

2012 ◽  
Vol 18 (4) ◽  
pp. 377-379 ◽  
Author(s):  
T.C.H. Hoogenboom ◽  
R.M.J. Van Beurden ◽  
B. Van Teylingen ◽  
B. Schenk ◽  
P.W.A. Willems

Time resolved whole brain CT angiography (4D-CTA) is a novel imaging technology providing information regarding blood flow. One of the factors that influence the diagnostic value of this examination is the temporal resolution, which is affected by the gantry rotation speed during acquisition and the reconstruction interval during post-processing. Post-processing determines the time spacing between two reconstructed volumes and, unlike rotation speed, does not affect radiation burden. The data sets of six patients who underwent a cranial 4D-CTA were used for this study. Raw data was acquired using a 320-slice scanner with a rotation speed of 2 Hz. The arterial to venous passage of an intravenous contrast bolus was captured during a 15 s continuous scan. The raw data was reconstructed using four different reconstruction-intervals: 0.2, 0.3, 0.5 and 1.0 s. The results were rated by two observers using a standardized score sheet. The appearance of each lesion was rated correctly in all readings. Scoring for quality of temporal resolution revealed a stepwise improvement from the 1.0 s interval to the 0.3 s interval, while no discernable improvement was noted between the 0.3 s and 0.2 s interval. An increase in temporal resolution may improve the diagnostic quality of cranial 4D-CTA. Using a rotation speed of 0.5 s, the optimal reconstruction interval appears to be 0.3 s, beyond which, changes can no longer be discerned.


Author(s):  
Eva-Maria Mandelkow ◽  
Eckhard Mandelkow ◽  
Joan Bordas

When a solution of microtubule protein is changed from non-polymerising to polymerising conditions (e.g. by temperature jump or mixing with GTP) there is a series of structural transitions preceding microtubule growth. These have been detected by time-resolved X-ray scattering using synchrotron radiation, and they may be classified into pre-nucleation and nucleation events. X-ray patterns are good indicators for the average behavior of the particles in solution, but they are difficult to interpret unless additional information on their structure is available. We therefore studied the assembly process by electron microscopy under conditions approaching those of the X-ray experiment. There are two difficulties in the EM approach: One is that the particles important for assembly are usually small and not very regular and therefore tend to be overlooked. Secondly EM specimens require low concentrations which favor disassembly of the particles one wants to observe since there is a dynamic equilibrium between polymers and subunits.


Author(s):  
T. Kizuka ◽  
N. Tanaka

Structure and stability of atomic clusters have been studied by time-resolved high-resolution electron microscopy (TRHREM). Typical examples are observations of structural fluctuation in gold (Au) clusters supported on silicon oxide films, graphtized carbon films and magnesium oxide (MgO) films. All the observations have been performed on the clusters consisted of single metal element. Structural stability of ceramics clusters, such as metal-oxide, metal-nitride and metal-carbide clusters, has not been observed by TRHREM although the clusters show anomalous structural and functional properties concerning to solid state physics and materials science.In the present study, the behavior of ceramic, magnesium oxide (MgO) clusters is for the first time observed by TRHREM at 1/60 s time resolution and at atomic resolution down to 0.2 nm.MgO and gold were subsequently deposited on sodium chloride (001) substrates. The specimens, single crystalline MgO films on which Au particles were dispersed were separated in distilled water and observed by using a 200-kV high-resolution electron microscope (JEOL, JEM2010) equipped with a high sensitive TV camera and a video tape recorder system.


Author(s):  
Eva-Maria Mandelkow ◽  
Ron Milligan

Microtubules form part of the cytoskeleton of eukaryotic cells. They are hollow libers of about 25 nm diameter made up of 13 protofilaments, each of which consists of a chain of heterodimers of α-and β-tubulin. Microtubules can be assembled in vitro at 37°C in the presence of GTP which is hydrolyzed during the reaction, and they are disassembled at 4°C. In contrast to most other polymers microtubules show the behavior of “dynamic instability”, i.e. they can switch between phases of growth and phases of shrinkage, even at an overall steady state [1]. In certain conditions an entire solution can be synchronized, leading to autonomous oscillations in the degree of assembly which can be observed by X-ray scattering (Fig. 1), light scattering, or electron microscopy [2-5]. In addition such solutions are capable of generating spontaneous spatial patterns [6].In an earlier study we have analyzed the structure of microtubules and their cold-induced disassembly by cryo-EM [7]. One result was that disassembly takes place by loss of protofilament fragments (tubulin oligomers) which fray apart at the microtubule ends. We also looked at microtubule oscillations by time-resolved X-ray scattering and proposed a reaction scheme [4] which involves a cyclic interconversion of tubulin, microtubules, and oligomers (Fig. 2). The present study was undertaken to answer two questions: (a) What is the nature of the oscillations as seen by time-resolved cryo-EM? (b) Do microtubules disassemble by fraying protofilament fragments during oscillations at 37°C?


2014 ◽  
Vol 134 (4) ◽  
pp. 515-520
Author(s):  
Jinfeng Yang ◽  
Yoichi Yoshida ◽  
Hiromi Shibata

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
C. Callenberg ◽  
A. Lyons ◽  
D. den Brok ◽  
A. Fatima ◽  
A. Turpin ◽  
...  

AbstractImaging across both the full transverse spatial and temporal dimensions of a scene with high precision in all three coordinates is key to applications ranging from LIDAR to fluorescence lifetime imaging. However, compromises that sacrifice, for example, spatial resolution at the expense of temporal resolution are often required, in particular when the full 3-dimensional data cube is required in short acquisition times. We introduce a sensor fusion approach that combines data having low-spatial resolution but high temporal precision gathered with a single-photon-avalanche-diode (SPAD) array with data that has high spatial but no temporal resolution, such as that acquired with a standard CMOS camera. Our method, based on blurring the image on the SPAD array and computational sensor fusion, reconstructs time-resolved images at significantly higher spatial resolution than the SPAD input, upsampling numerical data by a factor $$12 \times 12$$ 12 × 12 , and demonstrating up to $$4 \times 4$$ 4 × 4 upsampling of experimental data. We demonstrate the technique for both LIDAR applications and FLIM of fluorescent cancer cells. This technique paves the way to high spatial resolution SPAD imaging or, equivalently, FLIM imaging with conventional microscopes at frame rates accelerated by more than an order of magnitude.


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