Dynamic piecewise speckle temporal contrast imaging of microcirculation

2021 ◽  
Author(s):  
Jia Qin ◽  
Lin An
Author(s):  
Benjamin M. Siegel

The potential advantages of high voltage electron microscopy for extending the limits of resolution and contrast in imaging low contrast objects, such as biomolecular specimens, is very great. The results of computations will be presented showing that at accelerating voltages of 500-1000 kV it should be possible to achieve spacial resolutions of 1 to 1.5 Å and using phase contrast imaging achieve adequate image contrast to observe single atoms of low atomic number.The practical problems associated with the design and utilization of the high voltage instrument are, optimistically, within the range of competence of the state of the art. However, there are some extremely important and critical areas to be systematically investigated before we have achieved this competence. The basic electron optics of the column required is well understood, but before the full potential of an instrument capable of resolutions of better than 1.5 Å are realized some very careful development work will be required. Of great importance for the actual achievement of high resolution with a high voltage electron microscope is the fundamental limitation set by the characteristics of the high voltage electron beam that can be obtained from the accelerator column.


Author(s):  
Uwe Lücken ◽  
Michael Felsmann ◽  
Wim M. Busing ◽  
Frank de Jong

A new microscope for the study of life science specimen has been developed. Special attention has been given to the problems of unstained samples, cryo-specimens and x-ray analysis at low concentrations.A new objective lens with a Cs of 6.2 mm and a focal length of 5.9 mm for high-contrast imaging has been developed. The contrast of a TWIN lens (f = 2.8 mm, Cs = 2 mm) and the BioTWTN are compared at the level of mean and SD of slow scan CCD images. Figure 1a shows 500 +/- 150 and Fig. 1b only 500 +/- 40 counts/pixel. The contrast-forming mechanism for amplitude contrast is dependent on the wavelength, the objective aperture and the focal length. For similar image conditions (same voltage, same objective aperture) the BioTWIN shows more than double the contrast of the TWIN lens. For phasecontrast specimens (like thin frozen-hydrated films) the contrast at Scherzer focus is approximately proportional to the √ Cs.


Author(s):  
K. N. Colonna ◽  
G. Oliphant

Harmonious use of Z-contrast imaging and digital image processing as an analytical imaging tool was developed and demonstrated in studying the elemental constitution of human and maturing rabbit spermatozoa. Due to its analog origin (Fig. 1), the Z-contrast image offers information unique to the science of biological imaging. Despite the information and distinct advantages it offers, the potential of Z-contrast imaging is extremely limited without the application of techniques of digital image processing. For the first time in biological imaging, this study demonstrates the tremendous potential involved in the complementary use of Z-contrast imaging and digital image processing.Imaging in the Z-contrast mode is powerful for three distinct reasons, the first of which involves tissue preparation. It affords biologists the opportunity to visualize biological tissue without the use of heavy metal fixatives and stains. For years biologists have used heavy metal components to compensate for the limited electron scattering properties of biological tissue.


Author(s):  
Koenraad G F Janssens ◽  
Omer Van der Biest ◽  
Jan Vanhellemont ◽  
Herman E Maes ◽  
Robert Hull

There is a growing need for elastic strain characterization techniques with submicrometer resolution in several engineering technologies. In advanced material science and engineering the quantitative knowledge of elastic strain, e.g. at small particles or fibers in reinforced composite materials, can lead to a better understanding of the underlying physical mechanisms and thus to an optimization of material production processes. In advanced semiconductor processing and technology, the current size of micro-electronic devices requires an increasing effort in the analysis and characterization of localized strain. More than 30 years have passed since electron diffraction contrast imaging (EDCI) was used for the first time to analyse the local strain field in and around small coherent precipitates1. In later stages the same technique was used to identify straight dislocations by simulating the EDCI contrast resulting from the strain field of a dislocation and comparing it with experimental observations. Since then the technique was developed further by a small number of researchers, most of whom programmed their own dedicated algorithms to solve the problem of EDCI image simulation for the particular problem they were studying at the time.


Author(s):  
C. Barry Carter

This paper will review the current state of understanding of interface structure and highlight some of the future needs and problems which must be overcome. The study of this subject can be separated into three different topics: 1) the fundamental electron microscopy aspects, 2) material-specific features of the study and 3) the characteristics of the particular interfaces. The two topics which are relevant to most studies are the choice of imaging techniques and sample preparation. The techniques used to study interfaces in the TEM include high-resolution imaging, conventional diffraction-contrast imaging, and phase-contrast imaging (Fresnel fringe images, diffuse scattering). The material studied affects not only the characteristics of the interfaces (through changes in bonding, etc.) but also the method used for sample preparation which may in turn have a significant affect on the resulting image. Finally, the actual nature and geometry of the interface must be considered. For example, it has become increasingly clear that the plane of the interface is particularly important whenever at least one of the adjoining grains is crystalline.A particularly productive approach to the study of interfaces is to combine different imaging techniques as illustrated in the study of grain boundaries in alumina. In this case, the conventional imaging approach showed that most grain boundaries in ion-thinned samples are grooved at the grain boundary although the extent of this grooving clearly depends on the crystallography of the surface. The use of diffuse scattering (from amorphous regions) gives invaluable information here since it can be used to confirm directly that surface grooving does occur and that the grooves can fill with amorphous material during sample preparation (see Fig. 1). Extensive use of image simulation has shown that, although information concerning the interface can be obtained from Fresnel-fringe images, the introduction of artifacts through sample preparation cannot be lightly ignored. The Fresnel-fringe simulation has been carried out using a commercial multislice program (TEMPAS) which was intended for simulation of high-resolution images.


Author(s):  
J.-F. Revol ◽  
Y. Van Daele ◽  
F. Gaill

The only form of cellulose which could unequivocally be ascribed to the animal kingdom is the tunicin that occurs in the tests of the tunicates. Recently, high-resolution solid-state l3C NMR revealed that tunicin belongs to the Iβ form of cellulose as opposed to the Iα form found in Valonia and bacterial celluloses. The high perfection of the tunicin crystallites led us to study its crosssectional shape and to compare it with the shape of those in Valonia ventricosa (V.v.), the goal being to relate the cross-section of cellulose crystallites with the two allomorphs Iα and Iβ.In the present work the source of tunicin was the test of the ascidian Halocvnthia papillosa (H.p.). Diffraction contrast imaging in the bright field mode was applied on ultrathin sections of the V.v. cell wall and H.p. test with cellulose crystallites perpendicular to the plane of the sections. The electron microscope, a Philips 400T, was operated at 120 kV in a low intensity beam condition.


Author(s):  
D. R. Liu ◽  
D. B. Williams

The secondary electron imaging technique in a scanning electron microscope (SEM) has been used first by Millman et al. in 1987 to distinguish between the superconducting phase and the non-superconducting phase of the YBa2Cu3O7-x superconductors. They observed that, if the sample was cooled down below the transition temperature Tc and imaged with secondary electrons, some regions in the image would show dark contrast whereas others show bright contrast. In general, the contrast variation of a SEM image is the variation of the secondary electron yield over a specimen, which in turn results from the change of topography and conductivity over the specimen. Nevertheless, Millman et al. were able to demonstrate with their experimental results that the dominant contrast mechanism should be the conductivity variation and that the regions of dark contrast were the superconducting phase whereas the regions of bright contrast were the non-superconducting phase, because the latter was a poor conductor and consequently, the charge building-up resulted in high secondary electron emission. This observation has since aroused much interest amoung the people in electron microscopy and high Tc superconductivity. The present paper is the preliminary report of our attempt to carry out the secondary electron imaging of this material in a scanning transmission electron microscope (STEM) rather than in a SEM. The advantage of performing secondary electron imaging in a TEM is obvious that, in a TEM, the spatial resolution is higher and many more complementary techniques, e.g, diffraction contrast imaging, phase contrast imaging, electron diffraction and various microanalysis techniques, are available.


Author(s):  
Z. L. Wang ◽  
J. Bentley

The success of obtaining atomic-number-sensitive (Z-contrast) images in scanning transmission electron microscopy (STEM) has shown the feasibility of imaging composition changes at the atomic level. This type of image is formed by collecting the electrons scattered through large angles when a small probe scans across the specimen. The image contrast is determined by two scattering processes. One is the high angle elastic scattering from the nuclear sites,where ϕNe is the electron probe function centered at bp = (Xp, yp) after penetrating through the crystal; F denotes a Fourier transform operation; D is the detection function of the annular-dark-field (ADF) detector in reciprocal space u. The other process is thermal diffuse scattering (TDS), which is more important than the elastic contribution for specimens thicker than about 10 nm, and thus dominates the Z-contrast image. The TDS is an average “elastic” scattering of the electrons from crystal lattices of different thermal vibrational configurations,


Author(s):  
S. J. Pennycook

Using a high-angle annular detector on a high-resolution STEM it is possible to form incoherent images of a crystal lattice characterized by strong atomic number or Z contrast. Figure 1 shows an epitaxial Ge film on Si(100) grown by oxidation of Ge-implanted Si. The image was obtained using a VG Microscopes' HB501 STEM equipped with an ultrahigh resolution polepiece (Cs ∽1.2 mm, demonstrated probe FWHM intensity ∽0.22 nm). In both crystals the lattice is resolved but that of Ge shows much brighter allowing the interface to be located exactly and interface steps to be resolved (arrowed). The interface was indistinguishable in the phase-contrast STEM image from the same region, and even at higher resolution the location of the interface is complex. Figure 2 shows a thin region of an MBE-grown ultrathin super-lattice (Si8Ge2)100. The expected compositional modulation would show as one bright row of dots from the 2 Ge monolayers separated by 4 rows of lighter Si columns. The image shows clearly that strain-induced interdiffusion has occurred on the monolayer scale.


Author(s):  
S. J. Pennycook ◽  
P. D. Nellist ◽  
N. D. Browning ◽  
P. A. Langjahr ◽  
M. Rühle

The simultaneous use of Z-contrast imaging with parallel detection EELS in the STEM provides a powerful means for determining the atomic structure of grain boundaries. The incoherent Z-contrast image of the high atomic number columns can be directly inverted to their real space arrangement, without the use of preconceived structure models. Positions and intensities may be accurately quantified through a maximum entropy analysis. Light elements that are not visible in the Z-contrast image can be studied through EELS; their coordination polyhedra determined from the spectral fine structure. It even appears feasible to contemplate 3D structure refinement through multiple scattering calculations.The power of this approach is illustrated by the recent study of a series of SrTiC>3 bicrystals, which has provided significant insight into some of the basic issues of grain boundaries in ceramics. Figure 1 shows the structural units deduced from a set of 24°, 36° and 65° symmetric boundaries, and 24° and 45° asymmetric boundaries. It can be seen that apart from unit cells and fragments from the perfect crystal, only three units are needed to construct any arbitrary tilt boundary. For symmetric boundaries, only two units are required, each having the same Burgers, vector of a<100>. Both units are pentagons, on either the Sr or Ti sublattice, and both contain two columns of the other sublattice, imaging in positions too close for the atoms in each column to be coplanar. Each column was therefore assumed to be half full, with the pair forming a single zig-zag column. For asymmetric boundaries, crystal geometry requires two types of dislocations; the additional unit was found to have a Burgers’ vector of a<110>. Such a unit is a larger source of strain, and is especially important to the transport characteristics of cuprate superconductors. These zig-zag columns avoid the problem of like-ion repulsion; they have also been seen in TiO2 and YBa2Cu3O7-x and may be a general feature of ionic materials.


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