Complex wavefront reconstruction from multiple-image planes produced by a focus tunable lens

The two photon microscope provides optical sectioning of fluorescent specimens with a resolution comparable to that obtained in confocal microscopy (see refs 2,3). However, the excited volume in 2-photon microscopy is limited to the focal volume (unlike conventional fluorescence microscopy where excitation occurs throughout the specimen). This means that photodamage is limited to the plane of section being examined. Thus, the light emitted from each point in the specimen depends on the amount of fluorochrome present without the problem of prior illumination (of other planes within the specimen) reducing the photon yield so a better signal to noise ratio can be obtained when examination of multiple image planes is needed. Since 2-photon excitation spectra are wide and chromatic aberration is eliminated (because the emitted light does not have to be focused on a pinhole), it is possible to excite several fluorochromes simultaneously and map their positions with high accuracy.

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Using multiple image planes to achieve near-correct focus cues in a 3d display

Journal of Vision ◽

10.1167/4.8.463 ◽

2004 ◽

Vol 4 (8) ◽

pp. 463-463

Author(s):

S. J. Watt ◽

K. Akeley ◽

M. S. Banks

Keyword(s):

3D Display ◽

Multiple Image ◽

Focus Cues ◽

Image Planes

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DEVILS: a tool for the visualization of large datasets with a high dynamic range

F1000Research ◽

10.12688/f1000research.25447.2 ◽

2021 ◽

Vol 9 ◽

pp. 1380

Author(s):

Romain Guiet ◽

Olivier Burri ◽

Nicolas Chiaruttini ◽

Olivier Hagens ◽

Arne Seitz

Keyword(s):

Visual Inspection ◽

Dynamic Range ◽

Image Data ◽

High Dynamic Range ◽

Large Datasets ◽

Human Eye ◽

Multiple Image ◽

High Dynamic ◽

Image Planes ◽

Imagej Plugin

The number of grey values that can be displayed on monitors and be processed by the human eye is smaller than the dynamic range of image-based sensors. This makes the visualization of such data a challenge, especially with specimens where small dim structures are equally important as large bright ones, or whenever variations in intensity, such as non-homogeneous staining efficiencies or light depth penetration, becomes an issue. While simple intensity display mappings are easily possible, these fail to provide a one-shot observation that can display objects of varying intensities. In order to facilitate the visualization-based analysis of large volumetric datasets, we developed an easy-to-use ImageJ plugin enabling the compressed display of features within several magnitudes of intensities. The Display Enhancement for Visual Inspection of Large Stacks plugin (DEVILS) homogenizes the intensities by using a combination of local and global pixel operations to allow for high and low intensities to be visible simultaneously to the human eye. The plugin is based on a single, intuitively understandable parameter, features a preview mode, and uses parallelization to process multiple image planes. As output, the plugin is capable of producing a BigDataViewer-compatible dataset for fast visualization. We demonstrate the utility of the plugin for large volumetric image data.

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DEVILS: a tool for the visualization of large datasets with a high dynamic range

F1000Research ◽

10.12688/f1000research.25447.1 ◽

2020 ◽

Vol 9 ◽

pp. 1380

Author(s):

Romain Guiet ◽

Olivier Burri ◽

Nicolas Chiaruttini ◽

Olivier Hagens ◽

Arne Seitz

Keyword(s):

Visual Inspection ◽

Dynamic Range ◽

Image Data ◽

High Dynamic Range ◽

Large Datasets ◽

Human Eye ◽

Multiple Image ◽

High Dynamic ◽

Image Planes ◽

Imagej Plugin

The number of grey values that can be displayed on monitors and be processed by the human eye is smaller than the dynamic range of image-based sensors. This makes the visualization of such data a challenge, especially with specimens where small dim structures are equally important as large bright ones, or whenever variations in intensity, such as non-homogeneous staining efficiencies or light depth penetration, becomes an issue. While simple intensity display mappings are easily possible, these fail to provide a one-shot observation that can display objects of varying intensities. In order to facilitate the visualization-based analysis of large volumetric datasets, we developed an easy-to-use ImageJ plugin enabling the compressed display of features within several magnitudes of intensities. The Display Enhancement for Visual Inspection of Large Stacks plugin (DEVILS) homogenizes the intensities by using a combination of local and global pixel operations to allow for high and low intensities to be visible simultaneously to the human eye. The plugin is based on a single, intuitively understandable parameter, features a preview mode, and uses parallelization to process multiple image planes. As output, the plugin is capable of producing a BigDataViewer-compatible dataset for fast visualization. We demonstrate the utility of the plugin for large volumetric image data.

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Computer processing of high-resolution images of periodic specimens

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100141834 ◽

1986 ◽

Vol 44 ◽

pp. 2-5

Author(s):

W. Chiu ◽

M.F. Schmid ◽

T.-W. Jeng

Keyword(s):

High Resolution ◽

Fourier Transforms ◽

Complex Structure ◽

Distribution Functions ◽

Multiple Image ◽

Cryo Electron Microscopy ◽

Structure Factors ◽

Unit Cells ◽

High Resolution Images ◽

Phase Probability Distribution

Cryo-electron microscopy has been developed to the point where one can image thin protein crystals to 3.5 Å resolution. In our study of the crotoxin complex crystal, we can confirm this structural resolution from optical diffractograms of the low dose images. To retrieve high resolution phases from images, we have to include as many unit cells as possible in order to detect the weak signals in the Fourier transforms of the image. Hayward and Stroud proposed to superimpose multiple image areas by combining phase probability distribution functions for each reflection. The reliability of their phase determination was evaluated in terms of a crystallographic “figure of merit”. Grant and co-workers used a different procedure to enhance the signals from multiple image areas by vector summation of the complex structure factors in reciprocal space.

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Electron spectroscopic imaging (ESI) on multiphase polymer materials

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100153713 ◽

1989 ◽

Vol 47 ◽

pp. 348-349

Author(s):

H.-J. Cantow ◽

M. Kunz ◽

M. Möller

Keyword(s):

Energy Loss ◽

Melt Spinning ◽

Chromatic Aberration ◽

Element Distribution ◽

Polymer Materials ◽

Specific Energy Loss ◽

Transmission Electron ◽

Multicomponent Polymer ◽

Diffraction Patterns ◽

Image Planes

In transmission electron microscopy the natural contrast of polymers is very low. Thus the contrast has to be enhanced by staining with heavy metals. The resolution is limited by the size of the staining particles and by the fact that electrons with different energy are focused in different image planes due to the chromatic aberration of the magnetic lenses. The integration of an electron energy loss spectrometer into the optical coloumn of a transmission electron microscope offers the possibility to use monoenergetic electrons and to select electrons with a certain energy for imaging. Thus contrast and resolution are enhanced. By imaging only electrons with an element specific energy loss the element distribution in the sample can be obtained. In addition, elastic bright field images and diffraction patterns yield excellent resolution. Some applications of the method on multicomponent polymer materials are discussed.Bulk polymer samples were prepared by ultramicrotoming at room temperature or well below the glass transition temperature. Very thin films for the direct observation of the structure in semicrystalline polymers were obtained by melt-spinning. Specimens were examined with a ZEISS CEM 902 operated at 80 kV.

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Design of a Spectroscopic Low-Energy Emission Microscope

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100180549 ◽

1990 ◽

Vol 48 (1) ◽

pp. 358-359

Author(s):

Lee H. Veneklasen

Keyword(s):

Parallel Imaging ◽

Objective Lens ◽

Energy Window ◽

Flat Surfaces ◽

Beam Voltage ◽

New Instrument ◽

Real Time Image ◽

Image Planes ◽

Backscatter Diffraction ◽

Time Image

This paper discusses some of the unique aspects of a spectroscopic emission microscope now being tested in Clausthal. The instrument is designed for the direct parallel imaging of both elastic and inelastic electrons from flat surfaces. Elastic contrast modes of the familiar LEEM include large and small angle LEED, mirror microscopy, backscatter diffraction contrast (for imaging of surface structure), and phase contrast (for imaging of step dynamics)(1). Inelastic modes include topology sensitive secondary, and work function sensitive photoemission. Most important, the new instrument will also allow analytical imaging using characteristic Auger or soft X-ray emissions. The basic instrument has been described by Bauer and Telieps (2). This configuration has been redesigned to include an airlock, and a LaB6 gun, triple condensor lens, magnetic objective lens, a double focussing separator field, an imaging energy analyzer, and a real time image processor.Fig. 1 shows the new configuration. The basic beam voltage supply Vo = 20 KV, upon which separate supplies for the gun Vg, specimen Vs, lens electrode Vf, and analyzer bias Vb float. The incident energy at the sample can be varied from Vs = 0-1 KV for elastic imaging, or from Vg + Vs = (3 + Vs) KV for inelastic imaging. The image energy window Vs±V/2 may be varied without readjusting either the illumation, or imaging/analyzer optics. The diagram shows conjugate diffraction and image planes. The apertures defining incoming Humiliation and outgoing image angles are placed below the separator magnet to allow for their independent optimization. The instrument can illuminate and image 0.5-100 μm fields at 0-1 keV emission energies with an energy window down to 0.2 eV.

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