Tandem scanning reflected light microscopy: How it works and applications

1986 ◽  
Vol 44 ◽  
pp. 84-87
Author(s):  
Alan Boyde ◽  
Milan Hadravský ◽  
Mojmír Petran ◽  
Timothy F. Watson ◽  
Sheila J. Jones ◽  
...  
Keyword(s):  
Light Microscopy ◽  
Focal Plane ◽  
Central Symmetry ◽  
Single Line ◽  
Scanning Microscope ◽  
Reflected Light ◽  
Illuminating Light ◽  
Illuminating Beam

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.

GACH-prepared specimens for sem and confocal reflected light microscopy

1988 ◽  
Vol 46 ◽  
pp. 98-99
Author(s):  
R. E. Crang ◽  
B. Thompson
Keyword(s):  
Light Microscopy ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Scanning Microscope ◽  
Minimum Essential Medium ◽  
Culture Cells ◽  
Hindlimb Muscles ◽  
Tissue Culture Cells ◽  
Small Depth ◽  
Reflected Light

Biological specimens prepared for SEM are difficult to observe in detail by means of light microscopy. Conventional epi-illuminescence produces relatively poor images with reflected light due to poor contrast and multiple scattering of the light (1). However, we have found that mammalian tissue culture cells prepared for SEM by means of the co-polymerized glutaraldehyde-carbohydrazide (GACH) technique (2), can be alternately observed by means of SEM and reflected light microscopy if the light optical instrument is a confocal tandem scanning microscope (TSM). The images produced with TSM show resolution comparable to that of the SEM at magnifications ≤ 1,000 X. Furthermore, due to the extremely small depth of focus containing high contrast and brightness, optical sections can be produced which bear detail similar to that in conventional TEM observations.Fibroblasts and myotobes, from newborn rat hindlimb muscles were grown in Eagle's minimum essential medium with 10% fetal calf serum on collagen- coated glass cover slips for a period of 7 d (3).

Imaging sub-micron objects with light microscopy: Visualization of the T-4 bacteriophage

1989 ◽  
Vol 47 ◽  
pp. 834-835
Author(s):  
Malcolm Brown ◽  
Reynolds M. Delgado ◽  
Michael J. Fink
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Focal Plane ◽  
Phase Contrast ◽  
Dark Field ◽  
E Coli ◽  
Polystyrene Beads ◽  
Television Camera ◽  
Transmission Electron ◽  
Universal Microscope

While light microscopy has been used to image sub-micron objects, numerous problems with diffraction-limitations often preclude extraction of useful information. Using conventional dark-field and phase contrast light microscopy coupled with image processing, we have studied the following objects: (a) polystyrene beads (88nm, 264nm, and 557mn); (b) frustules of the diatom, Pleurosigma angulatum, and the T-4 bacteriophage attached to its host, E. coli or free in the medium. Equivalent images of the same areas of polystyrene beads and T-4 bacteriophages were produced using transmission electron microscopy.For light microscopy, we used a Zeiss universal microscope. For phase contrast observations a 100X Neofluar objective (N.A.=1.3) was applied. With dark-field, a 100X planachromat objective (N.A.=1.25) in combination with an ultra-condenser (N.A.=1.25) was employed. An intermediate magnifier (Optivar) was available to conveniently give magnification settings of 1.25, 1.6, and 2.0. The image was projected onto the back focal plane of a film or television camera with a Carl Zeiss Jena 18X Compens ocular.

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
Keyword(s):  
Electron Microscope ◽  
Light Microscopy ◽  
Carbon Films ◽  
Reflected Light

Hyperspectral reflected light microscopy of plasmonic Au/Ag alloy nanoparticles incubated as multiplex chromatic biomarkers with cancer cells

The Analyst ◽  
2014 ◽  
Vol 139 (20) ◽  
pp. 5247-5253 ◽  
Author(s):  
Sergiy Patskovsky ◽  
Eric Bergeron ◽  
David Rioux ◽  
Mikaël Simard ◽  
Michel Meunier
Keyword(s):  
Light Microscopy ◽  
Cancer Cells ◽  
Cancer Cell ◽  
Human Cancer ◽  
Spatial Localization ◽  
Alloy Nanoparticles ◽  
Human Cancer Cell ◽  
Plasmonic Nanoparticle ◽  
Reflected Light ◽  
Spectroscopic Identification

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.

REFLECTED LIGHT MICROSCOPY

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

Wood identification of charcoal with 3D-reflected light microscopy

IAWA Journal ◽  
2020 ◽  
Vol 41 (4) ◽  
pp. 478-489 ◽  
Author(s):  
Valentina Zemke ◽  
Volker Haag ◽  
Gerald Koch
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Light Microscopy ◽  
Field Emission ◽  
Polarized Light ◽  
Structural Features ◽  
Light Illumination ◽  
Reflected Light ◽  
Carbonized Wood ◽  
Scanning Electron

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.

Reflected-Light Microscopy

2019 ◽  
pp. 289-316
Keyword(s):  
Light Microscopy ◽  
Reflected Light

scholarly journals Curvature measurement with reflected-light microscopy and its application to fly facet lenses

1990 ◽  
Vol 158 (1) ◽  
pp. 87-93 ◽  
Author(s):  
D. G. Stavenga ◽  
H. L. Leertouwer
Keyword(s):  
Light Microscopy ◽  
Curvature Measurement ◽  
Reflected Light
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