A Correlative Approach to Colloidal Gold Labeling with Video-Enhanced Light Microscopy, Low-Voltage Scanning Electron Microscopy, and High-Voltage Electron Microscopy

Analysis of motility phenomena in a living cell observed with light microscopy can be significantly enriched by preparing a whole-mount of this cell for high voltage electron microscopy (to reveal the intracellular organization) and for low voltage scanning electron microscopy (to reveal the surface topography). In earlier studies, cell whole-mount prepration by chemical fixation and drying was adequate for studies of slow cellular motions at the subcellular level (e.g. receptor movements). Fast cellular motions analysed at the supramolecular level (e.g. transmitter release, cytoskeleton reorganization) required development of much faster cryo-immobilization methods. However, in studies of cells grown on grids, these freezing methods involved time consuming transfer of these cells , from an incubator to a freezer, making impossible fine correlations between images of a living cell and its cryo-whole-mount. To overcome this constraint for correlative microscopical studies of neoplastic cell motility, I designed an instrument consisting of a freezer attached to a light microscope and allowing cryoimmobilization within miliseconds after recording. The main objective of the current project was refinement of an instrument and improvement of appropriate specimen cryo-preparation techniques.

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Optimizing parameters for correlative immunogold localization by video-enhanced light microscopy, high-voltage transmission electron microscopy, and field emission scanning electron microscopy.

Journal of Histochemistry & Cytochemistry ◽

10.1177/38.12.2254644 ◽

1990 ◽

Vol 38 (12) ◽

pp. 1781-1785 ◽

Cited By ~ 14

Author(s):

S R Simmons ◽

J B Pawley ◽

R M Albrecht

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Transmission Electron Microscopy ◽

Light Microscopy ◽

High Voltage ◽

Low Voltage ◽

Immunogold Localization ◽

Transmission Electron ◽

High Voltage Transmission ◽

Scanning Electron

Correlative video-enhanced light microscopy, high-voltage transmission electron microscopy, and low-voltage high resolution scanning electron microscopy were used to examine the binding of colloidal gold-labeled fibrinogen to platelet surfaces. Optimal conditions for the detection of large (18 nm) and small (3 nm) gold particles are described.

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Novel Approaches to Low-Voltage Scanning Electron Microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100180586 ◽

1990 ◽

Vol 48 (1) ◽

pp. 366-367

Author(s):

Arthur V. Jones

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Low Voltage ◽

Electron Optics ◽

Current Interest ◽

New Concepts ◽

Opportune Time ◽

Novel Approaches ◽

Voltage Scanning ◽

Scanning Electron

In comparison with the developers of other forms of instrumentation, scanning electron microscope manufacturers are among the most conservative of people. New concepts usually must wait many years before being exploited commercially. The field emission gun, developed by Albert Crewe and his coworkers in 1968 is only now becoming widely available in commercial instruments, while the innovative lens designs of Mulvey are still waiting to be commercially exploited. The associated electronics is still in general based on operating procedures which have changed little since the original microscopes of Oatley and his co-workers.The current interest in low-voltage scanning electron microscopy will, if sub-nanometer resolution is to be obtained in a useable instrument, lead to fundamental changes in the design of the electron optics. Perhaps this is an opportune time to consider other fundamental changes in scanning electron microscopy instrumentation.

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Visualizing the surface morphology of non-fullerene acceptor blends by automated segmentation of spatially resolved electron spectra from ultra-low voltage scanning electron microscopy

10.29363/nanoge.hopv.2019.107 ◽

2019 ◽

Author(s):

Martin Pfannmöller ◽

Jochen Kammerer ◽

Rasmus Schröder ◽

Irene Wacker ◽

Riva Alkarsifi ◽

...

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Surface Morphology ◽

Low Voltage ◽

Automated Segmentation ◽

Electron Spectra ◽

Spatially Resolved ◽

Voltage Scanning ◽

Scanning Electron

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Low Voltage Scanning Electron Microscopy

Microscopy Today ◽

10.1017/s1551929500057813 ◽

2002 ◽

Vol 10 (2) ◽

pp. 22-23 ◽

Cited By ~ 2

Author(s):

David C Joy ◽

Dale E Newbury

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

High Spatial Resolution ◽

Low Voltage ◽

Incident Beam ◽

Operational Mode ◽

X Ray ◽

Rapid Fall ◽

Voltage Scanning ◽

Scanning Electron

Low Voltage Scanning Electron Microscopy (LVSEM), defined as operation in the energy range below 5 keV, has become perhaps the most important single operational mode of the SEM. This is because the LVSEM offers advantages in the imaging of surfaces, in the observation of poorly conducting and insulating materials, and for high spatial resolution X-ray microanalysis. These benefits all occur because a reduction in the energy Eo of the incident beam leads to a rapid fall in the range R of the electrons since R ∼k.E01.66. The reduction in the penetration of the beam has important consequences.

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