摘要:

AbstractThis paper proposes to evaluate, in tonsil lymphoid cells, the intracellular distribution of cyclin A, a protein involved in the cell cycle regulation. Serial sections of anti‐cyclin A labelled cells were optically acquired with a confocal laser scanning microscope. On these serial sections, three‐dimensional image analysis was applied, essentially by extending previous methods from the two‐dimensional to the three‐dimensional field. Sieving techniques derived from mathematical morphology were used to extract cluster‐ and spot‐shaped cyclin A structures. Subsequently, measurements were performed to estimate cell surface and volume, cyclin A contents, cyclin A patterns and cytoplasmic or nuclear cyclin A localization. Results were finally submitted to a principal component analysis which ordered the cells in such a way as to satisfy human visual perception. Cyclin A distribution appeared to vary greatly, and in a continuous way as no group was obviously shared. Some typical patterns, such as spotted cyclin A were observed. Confocal laser scanning microscopy and three‐dimensional analysis appeared as a relevant method to quantitatively investigate the subcellular distribution of proteins with respect to the fundamental three spatial dimens

ISSN:0966-9051    

DOI:10.1002/1361-6374(199406)2:2<69::AID-BIO1>3.0.CO;2-E

出版商:IOP Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

AbstractIn this paper two new recursive approaches are presented to correct for attenuation effects in fluorescent confocal microscopy, caused by absorption and scattering. The methods are based on a slice by slice processing of the three dimensional image using the result of the last processed layer to update the extinction information for correcting the next image plane. The first approach, Recursive Attenuation Correction using Directional Extinction Tracking (RAC‐DET), takes all directional extinction information into account, resulting in an accurate estimate of the original fluorescence distribution. By making additional assumptions about the imaging process a second approach taking into account only the extinction of the excitation is proposed: Recursive Attenuation Correction using Light front Tracking (RAC‐LT). Although correction accuracy is lower, computation times are greatly reduced such that an iterative scheme to estimate the extinction coefficient constant from the data becomes feasible for biological applicati

ISSN:0966-9051    

DOI:10.1002/1361-6374(199406)2:2<78::AID-BIO2>3.0.CO;2-D

出版商:IOP Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

AbstractThe complexity both of the measurements and of the specimens, when scanning tunnelling microscopy (STM) is applied in biology, requires a careful analysis of the data in order to avoid the classification of artifacts as peculiarities of the sample. The potential for determining the structures of biomolecules and their complexes emphasizes the relevance of developing procedures for STM image enhancement. Some software procedures have focused attention on the correction of geometrical distortion arising in STM images. It is our opinion that the application of such procedures may become very difficult when the real images are surfaces different from those used for calibration, i.e. graphite or mica. We have developed a procedure for reducing artifacts in STM and for providing image enhancement. It can be considered a noise filter that can restore the real images without knowledge of noise type and of noise ratio. We have termed it cross‐measurement procedure (CMP). CMP can be considered as a mixing of software and hardware procedures. In fact, it is based on two measurements of the same image performed by exchanging thexandyaxes and the origi

ISSN:0966-9051    

DOI:10.1002/1361-6374(199406)2:2<93::AID-BIO3>3.0.CO;2-N

出版商:IOP Publishing Ltd    

年代:1994

数据来源: WILEY

摘要:

AbstractWe have developed and tested an automated method for simultaneous 3‐D tracking of numerous, fluorescently‐tagged cells. The procedure uses multiple thresholding to segment individual cells at a starting timepoint, and then iteratively applies a template‐matching algorithm to locate a particular cell's position at subsequent timepoints. To speed up the method, we have developed a distributed implementation in which template matching is carried out in parallel on several different server machines. The distributed implementation showed a monotonic decrease in response time with increasing number of servers (up to 15 tested), demonstrating that the tracking algorithm is well suited to parallelization, and that nearly real‐time performance could be expected on a parallel processor. Of four different template matching statistics tested for 3‐D tracking of amoebae from the cellular slime mouldDictyostelium discoideum, we found that the automated procedure performed best when using a correlation statistic for matching. Using this statistic, the method achieved a 98.5% success rate in correctly identifying a cell from one time point to the next. This method is now being used regularly for 3‐D tracking of normal and mutant cells ofD. discoideum, and as such provides a means to quantify the motion of many cells within a three‐dimensiona

ISSN:0966-9051    

DOI:10.1002/1361-6374(199406)2:2<98::AID-BIO4>3.0.CO;2-5

出版商:IOP Publishing Ltd    

年代:1994

数据来源: WILEY

ISSN:0966-9051    

DOI:10.1002/1361-6374(199406)2:2<113::AID-BIO5>3.0.CO;2-L

出版商:IOP Publishing Ltd    

年代:1994

数据来源: WILEY

ISSN:0966-9051    

DOI:10.1002/1361-6374(199406)2:2<114::AID-BIO6>3.0.CO;2-F

出版商:IOP Publishing Ltd    

年代:1994

数据来源: WILEY