Fluorescence in the tandem scanning microscope

Boyde, A.; Jones, S. J.; Taylor, M.; Wolfe, L. A.; Watson, Timothy F.

doi:10.1111/j.1365-2818.1990.tb02945.x

Cited by 35 publications

(13 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Twenty freshly extracted, human lower third molar teeth were used in the study. This was considered essential, since the interface between restorative materials and tooth structure can be affected by post mortem changes in the tooth (Boyde et al, 1990). All teeth selected for the study were free of caries and surface defects, and had their pulpal contents intact.…”

Section: Methodsmentioning

confidence: 99%

Interfacial Characteristics of Resin-modified Glass-ionomer Materials: A Study on Fluid Permeability Using Confocal Fluorescence Microscopy

Sidhu

Watson

1998

J Dent Res

View full text Add to dashboard Cite

The tooth interface with resin-modified glass-ionomer cements (RM GICs) is poorly understood. This study examined the interface, especially with dentin. Cervical cavities in extracted teeth were restored with Fuji II LC, Vitremer, Photac-Fil, or a conventional GIC, Fuji Cap II. Fluorescent dye was placed in the pulp chambers for 3 hrs before the specimens were sectioned. Examination of the tooth/material interface with a confocal microscope showed that dye uptake by the restoration varied among materials. A "structureless", non-particulate, highly-stained layer of GIC was observed next to dentin in Fuji II LC. This layer varied in width, was prominent where the dentin tubules were cut "end-on" and in areas closer to the pulp, and was not seen adjacent to enamel. Vitremer showed minimal dye uptake, and the "structureless" layer was barely discernible. Photac-Fil showed more uniform uptake and absence of this layer. Cracking of enamel was also noted with these materials. The conventional GIC did not show any dye uptake, presence of a "structureless" layer, or enamel cracking. We elucidated the potential mechanisms involved in the formation of a "structureless" interfacial layer in Fuji II LC by studying the variables of cavity design, surface pre-treatment, water content of the tooth, time for it to develop, early finishing, and coating of the restoration. This layer, the "absorption layer", is probably related to water flux within the maturing cement, depending on environmental moisture changes and communication with the pulp in a wet tooth. The "micropermeability model" was useful in this study of the interfacial characteristics of RM GICs.

show abstract

Section: Methodsmentioning

confidence: 99%

Interfacial Characteristics of Resin-modified Glass-ionomer Materials: A Study on Fluid Permeability Using Confocal Fluorescence Microscopy

Sidhu

Watson

1998

J Dent Res

View full text Add to dashboard Cite

show abstract

“…For future use, further developments on the microscopy as well as the computational sector are expected: confocal microscopes operating under greater speed (Boyde et al, 1990;Goldstein et al, 1990) are being developed. Experiments on living cells, which have to be carried out at near video speed, require large data storage and processing capacities.…”

Section: Discussionmentioning

confidence: 99%

The three‐dimensional architecture of the mitotic spindle, analyzed by confocal fluorescence and electron microscopy

Merdes

Stelzer

Mey

1991

J. Elec. Microsc. Tech.

View full text Add to dashboard Cite

Fluorescence microscopy techniques have become important tools in mitosis research. The well-known disadvantages of fluorescence microscopy, rapid bleaching, phototoxicity and out-of-focus contributions blurring the in-focus image are obstacles which still need to be overcome. Confocal fluorescence microscopy has the potential to improve our capabilities of analyzing cells, because of its excellent depth-discrimination and image processing power. We have been using a confocal fluorescence microscope for the study of the mechanism of poleward chromosome movement, and report here 1) a cell preparation technique, which allows labeling of fixation sensitive spindle antigens with acceptable microtubule preservation; 2) the use of image processing methods to represent the spatial distribution of various labeled elements in pseudocolour; 3) a novel immunoelectron microscopic labeling method for microtubules, which allows the visualization of their distribution in semithin sections at low magnification; and 4) a first attempt to study microtubule dynamics with a confocal fluorescence microscope in living cells, microinjected with rhodamine labeled tubulin. Our experience indicates that confocal fluorescence microscopy provides real advantages for the study of spatial colocalization of antigens in the mitotic spindle. It does not, however, overcome the basic limits of resolution of the light microscope. Therefore, it has been necessary to use an electron microscopic method. Our preliminary results with living cells show that it is possible to visualize the entire microtubule network in stereo, but that the sensitivity of the instrument is still too low to perform dynamic time studies. It will be worthwhile to further develop this new type of optical instrumentation and explore its usefulness on both fixed and living cells.

show abstract

“…This is more so in formaldehyde fixed tissue, and may be dazzling in glutaraldehyde preserved material. If this is not enough, a general fluorescence is created by staining with brilliant sulphaflavine in ethanolic solution (Boyde et al, 1990): this dye is stable to electron beam irradiation and can also be used for cathodoluminescence (CL) mode imaging in the SEM (Boyde and Reid, 1984). Several strongly fluorescent dyes have the remarkable property of binding most strongly within calcified tissue matrix which is calcifying at the time of intra-vital administration (Frost, 1962;Hulth and Olerud, 1962;Rahn andPerren, 1970, 1971), although their distribution at lower concentrations is widespread in all hydrophobic compartments .…”

Section: Introductionmentioning

confidence: 99%

Combining confocal and BSE SEM imaging for bone block surfaces

Boyde

Lovicar²,

Zamecnik³

2005

eCM

View full text Add to dashboard Cite

The present report presents a method for the correlation of qualitative and quantitative BSE SEM imaging with confocal scanning light microscopy (CSLM) imaging modes applied to bone samples embedded in PMMA. The SEM has a proper digital scan generator: we leave the BSE image unchanged, and match the CSLM image to it, because the CSLM scan mechanism is not digital, though the signal is digitised. Our overlapping program uses a linear transformation matrix which projects one system to the other, calculated by finding three corresponding points in BSE and CSLM pictures.BSE images are empty where cells and osteoid are present. Fluorescence mode CSLM fills in these gaps. The combination images enhance our understanding of what is going on -and re-establish the need for good cellular preservation.

show abstract

Fluorescence in the tandem scanning microscope

Cited by 35 publications

References 19 publications

Interfacial Characteristics of Resin-modified Glass-ionomer Materials: A Study on Fluid Permeability Using Confocal Fluorescence Microscopy

Interfacial Characteristics of Resin-modified Glass-ionomer Materials: A Study on Fluid Permeability Using Confocal Fluorescence Microscopy

The three‐dimensional architecture of the mitotic spindle, analyzed by confocal fluorescence and electron microscopy

Combining confocal and BSE SEM imaging for bone block surfaces

Contact Info

Product

Resources

About