Imaging of tissue sections with very slow electrons

Frank, L.; Nebesářová, Jana; Vancová, Marie; Paták, Aleš; Müllerová, Ilona

doi:10.1016/j.ultramic.2014.10.009

Cited by 3 publications

(6 citation statements)

References 8 publications

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“…Unfortunately, only some details of the structure are highlighted in this way, mainly with staining. When using STEM equipped with a CL mode and decreasing the landing energy of electrons, we observe a dramatic increase in contrast even when no heavy metal-containing substances are used in preparation [16] (see Figure 16). An important finding made in a pioneering study [16] concerns changes in the embedding resin.…”

Section: Thin Sectionsmentioning

confidence: 97%

“…When using STEM equipped with a CL mode and decreasing the landing energy of electrons, we observe a dramatic increase in contrast even when no heavy metal-containing substances are used in preparation [16] (see Figure 16). An important finding made in a pioneering study [16] concerns changes in the embedding resin. It has been known that the resin is partially "radiation damaged" under electron bombardment with the consequence of the increased electron transmissivity of sections, but this increase was moderate only.…”

Section: Thin Sectionsmentioning

confidence: 97%

“…For this reason, the spatial density of the generation of the image information steeply increases so that, below 1 keV, we can get, for example, on tissue sections containing no agents with heavy metal salts not only very high contrast but also all structure details visualized, including those normally not highlighted with postfixation or staining media [16]. Naturally, the sections have to be thinner.…”

Section: Motivationmentioning

confidence: 99%

“…It has been known that the resin is partially "radiation damaged" under electron bombardment with the consequence of the increased electron transmissivity of sections, but this increase was moderate only. The rate of this effect was compared at various electron energies, and whereas 5 keV STEM increases the transmitted flux two times after a dose of about 5×10 -3 Ccm -2 , at 500 eV, the increase was 20× at a doubled dose [16]. The idea is that, under electron impact, the resin is partly depolymerized and monomers are then released from the surface; no losses in the observable structure details have been discovered after this "ultimate" preparation step performed in situ.…”

Section: Thin Sectionsmentioning

confidence: 99%

“…Figure16. Section of mouse heart muscle, not fixed with osmium tetroxide and not stained: 10 nm section imaged by conventional TEM at 80 keV (a), CL mode micrograph taken at 500 eV with a primary energy of 4.5 keV (b), and energy dependence of the contrast of this sample in the CL mode (c).…”

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confidence: 99%

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Scanning Electron Microscopy with a Retarded Primary Beam

Frank¹

2016

Modern Electron Microscopy in Physical and Life Sciences

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The general trend for reducing the energies of primary electrons in electron microscopy has been faced with a gradual deterioration of the image resolution. Biasing the sample to a high negative voltage and making the electrons arbitrarily slow solely on and inside the sample has shown itself to be far more feasible than originally expected. The fundamental aberration coefficients (spherical and chromatic) of a combination of an objective lens and an immersion electrostatic lens formed by the biased sample decrease with the decreasing landing energy of the electrons. As a result, the spot size in scanning systems may become nearly independent of the landing energy of the electrons. The requirements placed on samples are strict but feasible, and detection of signal electrons is greatly facilitated by the acceleration of both reflected and transmitted electrons in the field of the biased sample and their collimation toward the optical axis. The interaction of slow electrons is not only more intensive than that at standard energies but even scattering phenomena appear which are not otherwise observed. Several application examples are presented. The benefits of very low energy EM are still being uncovered after its having been in routine use for several years.

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Section: Thin Sectionsmentioning

confidence: 97%

Section: Thin Sectionsmentioning

confidence: 97%

Section: Motivationmentioning

confidence: 99%

Section: Thin Sectionsmentioning

confidence: 99%

mentioning

confidence: 99%

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Scanning Electron Microscopy with a Retarded Primary Beam

Frank¹

2016

Modern Electron Microscopy in Physical and Life Sciences

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Low Voltage STEM in the SEM

Brodusch

Demers

Gauvin

2017

Field Emission Scanning Electron Microscopy

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Quantification of STEM Images in High Resolution SEM for Segmented and Pixelated Detectors

et al. 2021

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The segmented semiconductor detectors for transmitted electrons in ultrahigh resolution scanning electron microscopes allow observing samples in various imaging modes. Typically, two standard modes of objective lens, with and without a magnetic field, differ by their resolution. If the beam deceleration mode is selected, then an electrostatic field around the sample is added. The trajectories of transmitted electrons are influenced by the fields below the sample. The goal of this paper is a quantification of measured images and theoretical study of the capability of the detector to collect signal electrons by its individual segments. Comparison of measured and ray-traced simulated data were difficult in the past. This motivated us to present a new method that enables better comparison of the two datasets at the cost of additional measurements, so-called calibration curves. Furthermore, we also analyze the measurements acquired using 2D pixel array detector (PAD) that provide a more detailed angular profile. We demonstrate that the radial profiles of STEM and/or 2D-PAD data are sensitive to material composition. Moreover, scattering processes are affected by thickness of the sample as well. Hence, comparing the two experimental and simulation data can help to estimate composition or the thickness of the sample.

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Imaging of tissue sections with very slow electrons

Cited by 3 publications

References 8 publications

Scanning Electron Microscopy with a Retarded Primary Beam

Scanning Electron Microscopy with a Retarded Primary Beam

Low Voltage STEM in the SEM

Quantification of STEM Images in High Resolution SEM for Segmented and Pixelated Detectors

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