Multi-dimensional and multi-signal approaches in scanning transmission electron microscopes

Colliex, C.; Brun, Nathalie; Gloter, Alexandre; Imhoff, D.; Kociak, Mathieu; March, Katia; Mory, C.; Stéphan, Odile; Tencé, Marcel; Walls, Michael

doi:10.1098/rsta.2009.0128

Cited by 20 publications

(10 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Currently, we are exploring the use of PINEM to image targeted sites of antibodies with immunolabeling (30) and the possibility of now varying a second time delay to examine dynamics with various specimen preparations, including cryogenic and even possibly biostructures at near-ambient conditions (31). It is also possible to vary the photon wavelength to map different dimensions, to further improve the spatial resolution by near-resonance confinement of the particle field, which is currently of 1-to 2-nm length scale (32), and the energy resolution for mapping all structures at once (33,34). The properties of the biological systems differ significantly from the inorganic materials previously studied (19), yet imaging with PINEM is still possible, suggesting possible extensions to a wide range of materials (ref.…”

Section: Resultsmentioning

confidence: 99%

Biological imaging with 4D ultrafast electron microscopy

Flannigan

Barwick

Zewail

2010

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Advances in the imaging of biological structures with transmission electron microscopy continue to reveal information at the nanometer length scale and below. The images obtained are static, i.e., time-averaged over seconds, and the weak contrast is usually enhanced through sophisticated specimen preparation techniques and/or improvements in electron optics and methodologies. Here we report the application of the technique of photon-induced near-field electron microscopy (PINEM) to imaging of biological specimens with femtosecond (fs) temporal resolution. In PINEM, the biological structure is exposed to single-electron packets and simultaneously irradiated with fs laser pulses that are coincident with the electron pulses in space and time. By electron energy-filtering those electrons that gained photon energies, the contrast is enhanced only at the surface of the structures involved. This method is demonstrated here in imaging of protein vesicles and whole cells of Escherichia coli, both are not absorbing the photon energy, and both are of low-Z contrast. It is also shown that the spatial location of contrast enhancement can be controlled via laser polarization, time resolution, and tomographic tilting. The high-magnification PINEM imaging provides the nanometer scale and the fs temporal resolution. The potential of applications is discussed and includes the study of antibodies and immunolabeling within the cell.evanescent | nanoscale | biostructure T he development and application of imaging techniques for the visualization of biological structures continues to advance our understanding of such systems. Various optical techniques have been developed to improve the spatial resolution beyond the diffraction limit (1-3) and to study, e.g., protein folding and adhesion complexes in living cells (4, 5). Though powerful, most optical methods rely on molecular components that emit light (fluoresce) and the spatial resolution cannot yet rival that of electron-based techniques that allow focusing down to the atomic scale (6). Force probe microscopies have been used to image surfaces of cells and porosomes with high enough spatial resolution (7), and pulsed X-ray sources, such as synchrotrons and free-electron lasers, hold promise for femtosecond (fs) diffraction studies of individual biological macromolecules (8). Biological imaging with electron microscopy goes back to the 1960s and has since then been advanced to enable structural mapping (9) of biological macromolecules and cells (10-12), including viruses and molecular machines such as the ribosome (6, 13).The visualization of biological structures with an electron microscope presents unique challenges, especially when considering the inherent weak contrast associated with such structures. This weak contrast, which is primarily because of the low-Z (atomic number) elemental composition of biological specimens and the need for thin samples, is often addressed by employing sophisticated specimen preparation techniques (e.g., ultrathin sectioning and staining) and by i...

show abstract

Section: Resultsmentioning

confidence: 99%

Biological imaging with 4D ultrafast electron microscopy

Flannigan

Barwick

Zewail

2010

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…More specifically, high-resolution scanning transmission electron microscopy (HRSTEM) techniques have shown unprecedented characterization possibilities due to the combination of high-resolution imaging, diffraction, and spectroscopy in a single experiment. [4] This characteristic allows the mapping of different electron-interaction events on the sample, which implies the possibility of probing features with accurate localization. In this sense, HRSTEM techniques point towards the quantitative characterization of the properties of materials with atomic resolution.…”

Section: Introductionmentioning

confidence: 99%

High‐Resolution Scanning Transmission Electron Microscopy (HRSTEM) Techniques: High‐Resolution Imaging and Spectroscopy Side by Side

et al. 2012

View full text Add to dashboard Cite

This work presents an overview of high-resolution scanning transmission electron microscopy (HRSTEM) techniques and exemplifies the novel quantitative characterization possibilities that have emerged from recent advances in these methods. The synergistic combination of atomic resolution imaging and spectroscopy provided by HRSTEM is highlighted as a unique feature that can provide a comprehensive analytical description of material properties at the nanoscale. State-of-the-art high-angle annular dark field and annular bright field examples are depicted as well as the use of X-ray energy-dispersive spectroscopy and electron energy-loss spectroscopy for probing samples properties at the atomic scale. In addition, promising techniques such as cathodoluminescence, confocal HRSTEM, and diffraction mapping are introduced. The presented examples and results indicate that HRSTEM-related techniques are fundamental tools for comprehensive assessment of properties at the atomic scale.

show abstract

“…1–3 These developments include efficient detectors, stable spectrometers, aberration correctors, as well as efficient algorithms for acquiring and processing the resulting spectra and images. Many of the same advances have also greatly enhanced our ability to perform EELS on biological systems, which however present some different challenges to the researcher.…”

Section: Introductionmentioning

confidence: 99%

Development of electron energy-loss spectroscopy in the biological sciences

Aronova

Leapman

2012

MRS Bull.

View full text Add to dashboard Cite

The high sensitivity of electron energy loss spectroscopy (EELS) for detecting light elements at the nanoscale makes it a valuable technique for application to biological systems. In particular, EELS provides quantitative information about elemental distributions within subcellular compartments, specific atoms bound to individual macromolecular assemblies, and the composition of bionanoparticles. The EELS data can be acquired either in the fixed beam energy-filtered transmission electron microscope (EFTEM) or in the scanning transmission electron microscope (STEM), and recent progress in the development of both approaches has greatly expanded the range of applications for EELS analysis. Near single atom sensitivity is now achievable for certain elements bound to isolated macromolecules, and it becomes possible to obtain three-dimensional compositional distributions from sectioned cells through EFTEM tomography.

show abstract

Multi-dimensional and multi-signal approaches in scanning transmission electron microscopes

Cited by 20 publications

References 25 publications

Biological imaging with 4D ultrafast electron microscopy

Biological imaging with 4D ultrafast electron microscopy

High‐Resolution Scanning Transmission Electron Microscopy (HRSTEM) Techniques: High‐Resolution Imaging and Spectroscopy Side by Side

Development of electron energy-loss spectroscopy in the biological sciences

Contact Info

Product

Resources

About