摘要:

A matching algorithm is proposed for aligning microscope images obtained using different modalities, making use of cross‐correlations of outputs from Prewitt's edge filter. Brightfield, phase contrast and differential interference contrast microscope images of algal and bacterial cells from an experimental, high‐rate algal pond are used for illustration. The information content of multimodal images is explored using principal components analysis and colour displays, and an image which represents optical thickness is constructed digita

ISSN:0022-2720    

DOI:10.1046/j.1365-2818.1996.91372.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

Confocal microscopes routinely produce three‐dimensional data sets. The visualization of these digital volumes is currently performed by one surface rendering or volume rendering approach. In this paper, we describe improvements developed in the field of volume rendering. We focused on three methods: parallelepiped face mapping: the rotation‐projection method (with or without stereoscopy, with different matters and transparencies); the voxel ray‐tracing method. We compared the possibilities of these different algorithms, in terms of quality of rendering, of computation load and as an essential aid to study the 3D organization of biological spec

ISSN:0022-2720    

DOI:10.1046/j.1365-2818.1996.117397.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

The bilateral imaging approach known from confocal applications operating in the line mode was used to realize real‐time two‐photon imaging. It is shown that the sectioning inherent to two‐photon imaging could be improved by the introduction of a confocal line aperture in the imaging path. Using a high‐power, low‐repetition‐rate amplified Ti:sapphire system, various biological objects were visualized including live

ISSN:0022-2720    

DOI:10.1046/j.1365-2818.1996.97379.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

Dark‐field imaging modes have attracted less interest in biological confocal microscopy than the extensive applications of immunofluorescence imaging. There are, however, certain biological and materials science applications where it is necessary to use a confocal imaging mode that is capable of partial or full rejection of light specularly reflected from coverslips, microscope slides or specimen surfaces. In this paper we present a simple modification of a commercial confocal microscope to incorporate dark‐field imaging. We discuss theoretical aspects of the resulting dark‐field imaging mode and also give experimental exa

ISSN:0022-2720    

DOI:10.1046/j.1365-2818.1996.127408.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

Cartilage, being highly aqueous, is difficult to preserve for electron microscopy without artefacts. Microwave‐enhanced fixation is suggested as a standard method for block samples of this material, with dimensions of up to 12 × 7 × 3 mm. Cartilage samples from the tibial plateau of adult rabbits were fixed by conventional, cryo‐ or microwave‐enhanced fixation. Constant or cyclical microwave irradiation of samples, immersed in fixatives, was carried out to varying final solution temperatures. Microwave‐enhanced fixation and staining is shown to be both rapid and reproducible, giving fine structural preservation. Below 323 K microwave fixation always gave excellent preservation of the fine structure within seconds. At higher temperatures thermal artefacts were introduced. In this study the microwave‐enhanced fixation is equal in quality to the best conventional immersion fixation and is nearly as fast as cryo‐preservation. It provides a standardized, reproducible fixation for morphological studies on cartilage

ISSN:0022-2720    

DOI:10.1046/j.1365-2818.1996.125406.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

A system of two opposite Everhart–Thornley detectors A and B has formerly been applied in conventional SEM for electron energies between 5 and 20 keV to separate material, topographic and other types of contrast by sum and difference signals. This technique can also be used successfully for low‐voltage scanning electron microscopy. The decreasing information depth with decreasing electron energy shows differences in the surface composition and contamination which cannot be observed beyond 5 keV. Also below 5 keV material and topographic contrast can be separated and increased by the A + B and A − B si

ISSN:0022-2720    

DOI:10.1046/j.1365-2818.1996.116396.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

Perfusion chambers employing laminar flow have dead volumes and unstirred layers which limit the minimum time required to effect a change in the local chemical environment of the sample. We have fabricated and tested a chamber capable of developing turbulent flow at reasonable flow rates of aqueous solutions. Transition to turbulence occurred at ≈1 mL s−1. To minimize dead space, a dual‐exit cross‐flow pattern was employed. The chamber was designed to mount on optical microscope stages for visual sample observation supplemented by a variety of techniques, such as fluorescence, light scattering and electrochemical monitoring. As indicated by fluorescence from a fluorescein‐labelled protein film adherent to the chamber wall, local pH changes were produced within 200 ms. Use of the chamber is illustrated by measurements of stopped‐flow kinetics in both calcium‐triggered cortical granule exocytosis and influenza virus haemagglutinin‐medi

ISSN:0022-2720    

DOI:10.1046/j.1365-2818.1996.103383.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY