Image Contrast

2003 ◽  
pp. 11-20
Author(s):  
Dominik Weishaupt ◽  
Victor D. Köchli ◽  
Borut Marincek
Keyword(s):  
Image Contrast

Progress In The Practical Application Of Automated Image Analysis To Tem Negatives

1977 ◽  
Vol 35 ◽  
pp. 214-217
Author(s):  
F. A. Heckman ◽  
E. Redman ◽  
J.E. Connolly
Keyword(s):  
Image Analysis ◽  
Image Quality ◽  
Exposure Time ◽  
Image Contrast ◽  
Automated Image Analysis ◽  
Practical Application ◽  
Factors Affecting ◽  
High Image Quality ◽  
Qualitative Evidence ◽  
Comprehensive Study

In our initial publication on this subject1) we reported results demonstrating that contrast is the most important factor in producing the high image quality required for reliable image analysis. We also listed the factors which enhance contrast in order of the experimentally determined magnitude of their effect. The two most powerful factors affecting image contrast attainable with sheet film are beam intensity and KV. At that time we had only qualitative evidence for the ranking of enhancing factors. Later we carried out the densitometric measurements which led to the results outlined below.Meaningful evaluations of the cause-effect relationships among the considerable number of variables in preparing EM negatives depend on doing things in a systematic way, varying only one parameter at a time. Unless otherwise noted, we adhered to the following procedure evolved during our comprehensive study:Philips EM-300; 30μ objective aperature; magnification 7000- 12000X, exposure time 1 second, anti-contamination device operating.

Observability of Very Thick Specimen by Using Transmission Scanning Electron Microscope

1971 ◽  
Vol 29 ◽  
pp. 26-27
Author(s):  
K. Shibatomi ◽  
T. Yamanoto ◽  
H. Koike
Keyword(s):  
Electron Microscope ◽  
Image Quality ◽  
Scanning Electron Microscope ◽  
Chromatic Aberration ◽  
Image Contrast ◽  
Objective Lens ◽  
Thick Specimen ◽  
Transmission Electron ◽  
Scanning Electron

In the observation of a thick specimen by means of a transmission electron microscope, the intensity of electrons passing through the objective lens aperture is greatly reduced. So that the image is almost invisible. In addition to this fact, it have been reported that a chromatic aberration causes the deterioration of the image contrast rather than that of the resolution. The scanning electron microscope is, however, capable of electrically amplifying the signal of the decreasing intensity, and also free from a chromatic aberration so that the deterioration of the image contrast due to the aberration can be prevented. The electrical improvement of the image quality can be carried out by using the fascionating features of the SEM, that is, the amplification of a weak in-put signal forming the image and the descriminating action of the heigh level signal of the background. This paper reports some of the experimental results about the thickness dependence of the observability and quality of the image in the case of the transmission SEM.

Single Atom Image Contrast in Fixed Beam Dark Field Electron Microscopy

1974 ◽  
Vol 32 ◽  
pp. 366-367
Author(s):  
Wah Chi
Keyword(s):  
Scattering Amplitude ◽  
Present Report ◽  
Dark Field ◽  
Image Contrast ◽  
Single Atom ◽  
Optical Theorem ◽  
Hartree Fock ◽  
Heavy Atoms ◽  
Beam Stop ◽  
Fixed Beam

Resolution and contrast are the important factors to determine the feasibility of imaging single heavy atoms on a thin substrate in an electron microscope. The present report compares the atom image characteristics in different modes of fixed beam dark field microscopy including the ideal beam stop (IBS), a wire beam stop (WBS), tilted illumination (Tl) and a displaced aperture (DA). Image contrast between one Hg and a column of linearly aligned carbon atoms (representing the substrate), are also discussed. The assumptions in the present calculations are perfectly coherent illumination, atom object is represented by spherically symmetric potential derived from Relativistic Hartree Fock Slater wave functions, phase grating approximation is used to evaluate the complex scattering amplitude, inelastic scattering is ignored, phase distortion is solely due to defocus and spherical abberation, and total elastic scattering cross section is evaluated by the Optical Theorem. The atom image intensities are presented in a Z-modulation display, and the details of calculation are described elsewhere.

Epoxy Resin Embedment

1968 ◽  
Vol 26 ◽  
pp. 100-101
Author(s):  
J. G. Adams ◽  
M. M. Campbell ◽  
H. Thomas ◽  
J. J. Ghldonl
Keyword(s):  
Electron Microscopy ◽  
Electron Beam ◽  
Epoxy Resin ◽  
Light Microscope ◽  
Epoxy Resins ◽  
Image Contrast ◽  
Tissue Preservation ◽  
Resin System ◽  
Excellent Quality

Since the introduction of epoxy resins as embedding material for electron microscopy, the list of new formulations and variations of widely accepted mixtures has grown rapidly. Described here is a resin system utilizing Maraglas 655, Dow D.E.R. 732, DDSA, and BDMA, which is a variation of the mixtures of Lockwood and Erlandson. In the development of the mixture, the Maraglas and the Dow resins were tested in 3 different volumetric proportions, 6:4, 7:3, and 8:2. Cutting qualities and characteristics of stability in the electron beam and image contrast were evaluated for these epoxy mixtures with anhydride (DDSA) to epoxy ratios of 0.4, 0.55, and 0.7. Each mixture was polymerized overnight at 60°C with 2% and 3% BDMA.Although the differences among the test resins were slight in terms of cutting ease, general tissue preservation, and stability in the beam, the 7:3 Maraglas to D.E.R. 732 ratio at an anhydride to epoxy ratio of 0.55 polymerized with 3% BDMA proved to be most consistent. The resulting plastic is relatively hard and somewhat brittle which necessitates trimming and facing the block slowly and cautiously to avoid chipping. Sections up to about 2 microns in thickness can be cut and stained with any of several light microscope stains and excellent quality light photomicrographs can be taken of such sections (Fig. 1).

Scanning Transmission Microscopy Applied to Thick Specimen

1972 ◽  
Vol 30 ◽  
pp. 452-453
Author(s):  
H. Koike ◽  
T. Matsuo ◽  
K. Ueno ◽  
M. Suzuki
Keyword(s):  
Psoas Muscle ◽  
Image Intensifier ◽  
Chromatic Aberration ◽  
Image Contrast ◽  
Transmission Microscopy ◽  
Crystalline Materials ◽  
Thick Specimen ◽  
Transmission Electron ◽  
Scanning Transmission ◽  
Electron Microscope Image

Since the identification of single atoms was achieved by Crewe et al, scanning transmission microscopy has been put into pratical use. Recently they applied this method to the quantitative mass analysis of DNA.As pointed out previously the chromatic aberration which decreases the image contrast and quality, does not affect a scanning transmission image as it does a conventional transmission electron microscope image. Thus, the STEM method is advantageous for thick specimen. Further this method employs a high sensitive photomultiplier tube which also functions as an image intensifier. This detection method is effective for the observation of living specimens or easily damaged specimens. In this respect the scanning transmission microscope with high accelerating voltage is necessary.Since Uyeda's experiments of crystalline materials, many workers have been discussed how thick specimens can be observed by CTEM. With biological specimens, R. Szirmae reported on the decrease in the image contrast of rabbit psoas muscle sections at various accelerating voltages and specimen thicknesses.

Analysis of Contrast Reversal in SEM Images

1972 ◽  
Vol 30 ◽  
pp. 398-399
Author(s):  
M. D. Coutts ◽  
E. R. Levin
Keyword(s):  
Incident Beam ◽  
Normal Incidence ◽  
Beam Current ◽  
Secondary Emission ◽  
Image Contrast ◽  
Sem Images ◽  
Secondary Electrons ◽  
Image Position

On tilting samples in an SEM, the image contrast between two elements, x and y often decreases to zero at θε, which we call the no-contrast angle. At angles above θε the contrast is reversed, θ being the angle between the specimen normal and the incident beam. The available contrast between two elements, x and y, in the SEM can be defined as,(1)where ix and iy are the total number of reflected and secondary electrons, leaving x and y respectively. It can easily be shown that for the element x,(2)where ib is the beam current, isp the specimen absorbed current, δo the secondary emission at normal incidence, k is a constant, and m the reflected electron coefficient.

Stain contamination and embedding in electron microscopy

1986 ◽  
Vol 44 ◽  
pp. 50-53
Author(s):  
Hilton H. Mollenhauer
Keyword(s):  
Electron Microscopy ◽  
Information Retrieval ◽  
Epoxy Resins ◽  
Image Contrast ◽  
Sample Preservation ◽  
Factors Associated ◽  
Amount Of Information ◽  
Electron Microscopical ◽  
Contrast Information

Many factors (e.g., resolution of microscope, type of tissue, and preparation of sample) affect electron microscopical images and alter the amount of information that can be retrieved from a specimen. Of interest in this report are those factors associated with the evaluation of epoxy embedded tissues. In this context, informational retrieval is dependant, in part, on the ability to “see” sample detail (e.g., contrast) and, in part, on tue quality of sample preservation. Two aspects of this problem will be discussed: 1) epoxy resins and their effect on image contrast, information retrieval, and sample preservation; and 2) the interaction between some stains commonly used for enhancing contrast and information retrieval.

Computer-Assisted High-Re Solution TEM

1990 ◽  
Vol 48 (1) ◽  
pp. 162-163
Author(s):  
F.A. Ponce ◽  
H. Hikashi
Keyword(s):  
Signal To Noise Ratio ◽  
Accurate Determination ◽  
Computer Software ◽  
Video Camera ◽  
Image Contrast ◽  
Objective Lens ◽  
Computer Assisted ◽  
Optical Parameters ◽  
Astigmatism Correction

The determination of the atomic positions from HRTEM micrographs is only possible if the optical parameters are known to a certain accuracy, and reliable through-focus series are available to match the experimental images with calculated images of possible atomic models. The main limitation in interpreting images at the atomic level is the knowledge of the optical parameters such as beam alignment, astigmatism correction and defocus value. Under ordinary conditions, the uncertainty in these values is sufficiently large to prevent the accurate determination of the atomic positions. Therefore, in order to achieve the resolution power of the microscope (under 0.2nm) it is necessary to take extraordinary measures. The use of on line computers has been proposed [e.g.: 2-5] and used with certain amount of success.We have built a system that can perform operations in the range of one frame stored and analyzed per second. A schematic diagram of the system is shown in figure 1. A JEOL 4000EX microscope equipped with an external computer interface is directly linked to a SUN-3 computer. All electrical parameters in the microscope can be changed via this interface by the use of a set of commands. The image is received from a video camera. A commercial image processor improves the signal-to-noise ratio by recursively averaging with a time constant, usually set at 0.25 sec. The computer software is based on a multi-window system and is entirely mouse-driven. All operations can be performed by clicking the mouse on the appropiate windows and buttons. This capability leads to extreme friendliness, ease of operation, and high operator speeds. Image analysis can be done in various ways. Here, we have measured the image contrast and used it to optimize certain parameters. The system is designed to have instant access to: (a) x- and y- alignment coils, (b) x- and y- astigmatism correction coils, and (c) objective lens current. The algorithm is shown in figure 2. Figure 3 shows an example taken from a thin CdTe crystal. The image contrast is displayed for changing objective lens current (defocus value). The display is calibrated in angstroms. Images are stored on the disk and are accessible by clicking the data points in the graph. Some of the frame-store images are displayed in Fig. 4.

Scattering of Electrons at High - Pressure and Restoration of Image Contrast in High Pressure Scanning Electron Microscopy

1990 ◽  
Vol 48 (1) ◽  
pp. 384-385
Author(s):  
J. S. Shah ◽  
R. Durkin ◽  
A. N. Farley
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
High Pressure ◽  
Cross Sections ◽  
Single Scattering ◽  
Image Contrast ◽  
Experimental Scheme ◽  
Medium Resolution ◽  
Scattering Approximation ◽  
Scanning Electron

It is now possible to perform High Pressure Scanning Electron Microscopy (HPSEM) in the range 10 to 2000 Pa. Here the effect of scattering on resolution has been evaluated by calculating the profile of the beam in high pressure and assessing its effect on the image contrast . An experimental scheme is presented to show that the effect of the primary beam ionization is to reduce image contrast but this effect can be eliminated by a novel use of specimen current detection in the presence of an electric field. The mechanism of image enhancement is discussed in terms of collection of additional carriers generated by the emissive components.High Pressure SEM (HPSEM) instrumentation is establishing itself as commercially viable. There are now a number of manufacturers, such as JEOL, ABT, ESCAN, DEBEN RESEARCH, selling microscopes and accessories for HPSEM. This is because high pressure techniques have begun to yield high quality micrographs at medium resolution.To study the effect of scattering on the incident electron beam, its profile - in a high pressure environment - was evaluated by calculating the elastic and inelastic scattering cross sections for nitrogen in the energy range 5-25 keV. To assess the effect of the scattered beam on the image contrast, the modification of a sharp step contrast function due to scattering was calculated by single scattering approximation and experimentally confirmed for a 20kV accelerated beam.

Observations of Frozen-Hydrated Microtubules

1990 ◽  
Vol 48 (1) ◽  
pp. 504-505
Author(s):  
D. Chrétien ◽  
D. Job ◽  
R.H. Wade
Keyword(s):  
Fourier Transforms ◽  
Structural Complexity ◽  
Image Contrast ◽  
Phase Behaviour ◽  
Experimental Conditions ◽  
Thin Sections ◽  
Surface Lattice ◽  
Cilia And Flagella ◽  
Outstanding Question

Microtubules are filamentary structures found in the cytoplasm of eukaryotic cells, where, together with actin and intermediate filaments, they form the components of the cytoskeleton. They have many functions and show various levels of structural complexity as witnessed by the singlet, doublet and triplet structures involved in the architecture of centrioles, basal bodies, cilia and flagella. The accepted microtubule model consists of a 25 nm diameter hollow tube with a wall made up of 13 paraxial protofilaments (pf). Each pf is a string of aligned tubulin dimers. Some results have suggested that the pfs follow a superhelix. To understand how microtubules function in the cell an accurate model of the surface lattice is one of the requirements. For example the 9x2 architecture of the axoneme will depend on the organisation of its component microtubules. We should also note that microtubules with different numbers of pfs have been observed in thin sections of cellular and of in-vitro material. An outstanding question is how does the surface lattice adjust to these different pf numbers?We have been using cryo-electron microscopy of frozen-hydrated samples to study in-vitro assembled microtubules. The experimental conditions are described in detail in this reference. The results obtained in conjunction with thin sections of similar specimens and with axoneme outer doublet fragments have already allowed us to characterise the image contrast of 13, 14 and 15 pf microtubules on the basis of the measured image widths, of the the image contrast symmetry and of the amplitude and phase behaviour along the equator in the computed Fourier transforms. The contrast variations along individual microtubule images can be interpreted in terms of the geometry of the microtubule surface lattice. We can extend these results and make some reasonable predictions about the probable surface lattices in the case of other pf numbers, see Table 1. Figure 1 shows observed images with which these predictions can be compared.

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