Defects of Planarity of Carbon Films Supported on Electron Microscope Grids Revealed by Reflected Light Microscopy

1994 ◽  
Vol 112 (3) ◽  
pp. 252-258 ◽  
Author(s):  
Marc Schmutz ◽  
Jacques Lang ◽  
Sabine Graff ◽  
Alain Brisson
Author(s):  
C. T. Nightingale ◽  
S. E. Summers ◽  
T. P. Turnbull

The ease of operation of the scanning electron microscope has insured its wide application in medicine and industry. The micrographs are pictorial representations of surface topography obtained directly from the specimen. The need to replicate is eliminated. The great depth of field and the high resolving power provide far more information than light microscopy.


Author(s):  
C. C. Clawson ◽  
L. W. Anderson ◽  
R. A. Good

Investigations which require electron microscope examination of a few specific areas of non-homogeneous tissues make random sampling of small blocks an inefficient and unrewarding procedure. Therefore, several investigators have devised methods which allow obtaining sample blocks for electron microscopy from region of tissue previously identified by light microscopy of present here techniques which make possible: 1) sampling tissue for electron microscopy from selected areas previously identified by light microscopy of relatively large pieces of tissue; 2) dehydration and embedding large numbers of individually identified blocks while keeping each one separate; 3) a new method of maintaining specific orientation of blocks during embedding; 4) special light microscopic staining or fluorescent procedures and electron microscopy on immediately adjacent small areas of tissue.


Author(s):  
J. H. Luft

Ruthenium red is one of the few completely inorganic dyes used to stain tissues for light microscopy. This novelty is enhanced by ignorance regarding its staining mechanism. However, its continued usefulness in botany for demonstrating pectic substances attests to selectivity of some sort. Whether understood or not, histochemists continue to be grateful for small favors.Ruthenium red can also be used with the electron microscope. If single cells are exposed to ruthenium red solution, sufficient mass can be bound to produce observable density in the electron microscope. Generally, this effect is not useful with solid tissues because the contrast is wasted on the damaged cells at the block surface, with little dye diffusing more than 25-50 μ into the interior. Although these traces of ruthenium red which penetrate between and around cells are visible in the light microscope, they produce negligible contrast in the electron microscope. However, its presence can be amplified by a reaction with osmium tetroxide, probably catalytically, to be easily visible by EM. Now the density is clearly seen to be extracellular and closely associated with collagen fibers (Fig. 1).


Author(s):  
M. K. Lamvik

When observing small objects such as cellular organelles by scanning electron microscopy, it is often valuable to use the techniques of transmission electron microscopy. The common practice of mounting and coating for SEM may not always be necessary. These possibilities are illustrated using vertebrate skeletal muscle myofibrils.Micrographs for this study were made using a Hitachi HFS-2 scanning electron microscope, with photographic recording usually done at 60 seconds per frame. The instrument was operated at 25 kV, with a specimen chamber vacuum usually better than 10-7 torr. Myofibrils were obtained from rabbit back muscle using the method of Zak et al. To show the component filaments of this contractile organelle, the myofibrils were partially disrupted by agitation in a relaxing medium. A brief centrifugation was done to clear the solution of most of the undisrupted myofibrils before a drop was placed on the grid. Standard 3 mm transmission electron microscope grids covered with thin carbon films were used in this study.


Author(s):  
Alan Boyde ◽  
Milan Hadravský ◽  
Mojmír Petran ◽  
Timothy F. Watson ◽  
Sheila J. Jones ◽  
...  

The principles of tandem scanning reflected light microscopy and the design of recent instruments are fully described elsewhere and here only briefly. The illuminating light is intercepted by a rotating aperture disc which lies in the intermediate focal plane of a standard LM objective. This device provides an array of separate scanning beams which light up corresponding patches in the plane of focus more intensely than out of focus layers. Reflected light from these patches is imaged on to a matching array of apertures on the opposite side of the same aperture disc and which are scanning in the focal plane of the eyepiece. An arrangement of mirrors converts the central symmetry of the disc into congruency, so that the array of apertures which chop the illuminating beam is identical with the array on the observation side. Thus both illumination and “detection” are scanned in tandem, giving rise to the name Tandem Scanning Microscope (TSM). The apertures are arranged on Archimedean spirals: each opposed pair scans a single line in the image.


Author(s):  
George Guthrie ◽  
David Veblen

The nature of a geologic fluid can often be inferred from fluid-filled cavities (generally <100 μm in size) that are trapped during the growth of a mineral. A variety of techniques enables the fluids and daughter crystals (any solid precipitated from the trapped fluid) to be identified from cavities greater than a few micrometers. Many minerals, however, contain fluid inclusions smaller than a micrometer. Though inclusions this small are difficult or impossible to study by conventional techniques, they are ideally suited for study by analytical/ transmission electron microscopy (A/TEM) and electron diffraction. We have used this technique to study fluid inclusions and daughter crystals in diamond and feldspar.Inclusion-rich samples of diamond and feldspar were ion-thinned to electron transparency and examined with a Philips 420T electron microscope (120 keV) equipped with an EDAX beryllium-windowed energy dispersive spectrometer. Thin edges of the sample were perforated in areas that appeared in light microscopy to be populated densely with inclusions. In a few cases, the perforations were bound polygonal sides to which crystals (structurally and compositionally different from the host mineral) were attached (Figure 1).


The Analyst ◽  
2014 ◽  
Vol 139 (20) ◽  
pp. 5247-5253 ◽  
Author(s):  
Sergiy Patskovsky ◽  
Eric Bergeron ◽  
David Rioux ◽  
Mikaël Simard ◽  
Michel Meunier

We report a hyperspectral reflected light microscopy system for plasmonic nanoparticle (NP) imaging, and compare with a conventional darkfield method for spatial localization and spectroscopic identification of single Au, Ag and Au/Ag alloy NPs incubated with fixed human cancer cell preparations.


Development ◽  
1984 ◽  
Vol 80 (1) ◽  
pp. 321-330
Author(s):  
Jonathan M. W. Slack

The development of ectoderm isolated from the animal pole of axolotl gastrulae is monitored by light microscopy, electron microscopy and analysis of newly synthesized proteins, glycoproteins and glycolipids. When control embryos are undergoing neurulation it is shown that the explants autonomously begin to express epidermal markers and do not express mesodermal markers. However the results suggest that not all the cells become epidermal and electron microscope examination shows that only the outer layer does so, the inner cells remaining undifferentiated.


1966 ◽  
pp. 1-18
Author(s):  
R.E. SMALLMAN ◽  
K.H.G. ASHBEE

IAWA Journal ◽  
2020 ◽  
Vol 41 (4) ◽  
pp. 478-489 ◽  
Author(s):  
Valentina Zemke ◽  
Volker Haag ◽  
Gerald Koch

Abstract The present study focusses on the application of 3D-reflected light microscopy (3D-RLM) for the wood anatomical identification of charcoal specimens produced from domestic and tropical timbers. This special microscopic technique offers a detailed investigation of anatomical features in charcoal directly compared with the quality of field emission scanning electron microscopy (FESEM). The advantages of using the 3D-RLM technology are that fresh fracture planes of charcoal can be directly observed under the microscope without further preparation or surface treatment. Furthermore, the 3D-technique with integrated polarized light illumination creates high-contrast images of uneven and black charcoal surfaces. Important diagnostic structural features such as septate fibres and intercellular canals can be clearly detected and intervessel pits are directly measured. The comparison of the microscopic analyses reveals that 3D-reflected light microscopy (3D-RLM) provides an effective alternative technique to conventional field emission scanning electron microscopy for the identification of carbonized wood.


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