摘要:

AbstractA stand‐alone atomic force microscope (AFM) has been developed, which features a large scan area and which allows operation under liquid. This system was combined with a confocal laser scanning microscope (CLSM). Information about cell structures, obtained by CLSM, can be complemented with images of the cell surface obtained with the AFM. This is illustrated by studying the pseudopodia of cells from a human cell line (K562‐cells, predecessor of erythroblasts) and the cytoskeleton of monkey kidney cells (in air and under liquid), both stained with F‐actin‐specific fluorescent probes. Images of the cytoskeleton during the cytotoxic interaction between a natural killer and a K562 target cell are presented. Our results show that combination of these techniques can provide new information about cells and cellular str

ISSN:0966-9051    

DOI:10.1002/1361-6374(199306)1:2<63::AID-BIO1>3.0.CO;2-Y

出版商:IOP Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

AbstractA new modular transmission and fluorescence illumination system has been designed and built. The approach utilizes fiber optics to scramble the spatial intensity variations and digital monitoring of light intensity to allow accurate correction for inherent temporal instability. The illumination system has been characterized by a variety of methods and is shown to have an evenly filled objective back focal plane for maximal resolution in all three axes. The use of this illumination system in conjunction with a scientific grade CCD camera and pixel‐by‐pixel correction makes possible data of such high quality as to be only limited by the photon counting statistics. This illumination approach has proven to be particularly important for three‐dimensional imaging coupled with image processing to remove out‐of‐focus in

ISSN:0966-9051    

DOI:10.1002/1361-6374(199306)1:2<71::AID-BIO2>3.0.CO;2-#

出版商:IOP Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

AbstractTwo biological specimens typical of 3‐D samples studied by confocal microscopy were imaged at matched focal planes in conventional and confocal microscopes. A comparison of the effectiveness of optical sectioning was made between confocal microscopy and wide‐field microscopy with computer deconvolution (using commercially available image restoration software). A simple algorithm for improving the depth discrimination of a confocal microscope is described and demonstra

ISSN:0966-9051    

DOI:10.1002/1361-6374(199306)1:2<82::AID-BIO3>3.0.CO;2-T

出版商:IOP Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

AbstractThe three‐dimensional positions of two centromeric DNA probes specific for chromosomes 7 and 15 were determined using optical sectioning by multi‐wavelength confocal laser scanning microscopy of human interphase fibroblasts. In addition to the fluorescencein situhybridization (FISH) signals, the immunofluorescence of nuclear lamin was used to delineate the nuclear boundary. The coordinates of thein situprobes and the centre of the nucleus were determined for 38 nuclei from which the vectors and distances between homologous and heterologous centromeres and from each centromere to the nuclear centre were computed. The distributions of the calculated distances were analysed statistically and compared with those predicted for a random distribution. It was found that both chromosome loci are non‐randomly distributed in the nucleus and that centromere 15, compared with centromere 7, is significantly closer to the centre of the nucleus. We established that quantitative three‐dimensional (3‐D) measurements of multiple targets are feasible although the experimentally and computationally intensive methods limit the attainable dataset to a relatively small number of nuclei. Therefore, the results were compared with those acquired for a larger number of similar cells in a simpler two‐dimensional (2‐D) analysis carried out in a co

ISSN:0966-9051    

DOI:10.1002/1361-6374(199306)1:2<96::AID-BIO4>3.0.CO;2-D

出版商:IOP Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

AbstractTwo‐dimensional fluorescence images were used to determine the localization ofin situhybridization signals from two centromeric DNA probes within human fibroblasts as a function of the cell cycle. The probes were specific for the centromeres of chromosomes 7 and 15. A high resolution CCD camera combined with image processing software allowed a semi‐automated quantitation of the data, which were analysed statistically. It was found that the distances of centromere 15 to the nuclear membrane were significantly greater than those of centromere 7, the distribution of which was random in cycling cells. The distances between the homolog 15 centromeres were smaller than between the homolog 7 centromeres. In addition, both centromeric probes showed the largest deviation from a random orientation in theG0resting state of the cell cycle. The more central location of centromere 15 may reflect the localization of the nucleolus organizing region of this chromosome and the corresponding location of nucleoli in human fibroblasts. The methods used to acquire and process the data are well suited to the semi‐automated analysis of fluorescencein situhybridization (FISH) signals for the cytogenetic analysis of normal and pathologic t

ISSN:0966-9051    

DOI:10.1002/1361-6374(199306)1:2<107::AID-BIO5>3.0.CO;2-G

出版商:IOP Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

AbstractThe registration of image volumes has been described in several publications for different imaging modalities, such as MRI, PET, SPECT and CT. However, for the 3‐D alignment of histology and MRI investigations concerning the influence of local nonlinear distortions are required. A non‐iterative and noninteractive least‐squares 3‐D affine transformation with trilinear interpolation in conjunction with a correlation‐based 3‐D searching algorithm was developed in order to estimate the influence of local deformations and the number of reference points with respect to the best transformation parameters. The application of the implemented software is demonstrated with data from human brains, which were visualized using magnetic resonance imaging and subsequently in histological sections. The procedures are presented with a discussion of quantitative error analysis for assessing the usefulness of the transformation under real conditions, e.g., the projections of the surface reconstructions in three orthogonal directions revealed a misalignment of 1.9, 2.9 and 3.1%, r

ISSN:0966-9051    

DOI:10.1002/1361-6374(199306)1:2<119::AID-BIO6>3.0.CO;2-6

出版商:IOP Publishing Ltd    

年代:1993

数据来源: WILEY