Combined confocal and wide-field high-resolution cytometry of FISH-stained cells
|Year of publication
|Article in Periodical
|Magazine / Source
|MU Faculty or unit
|Use of computers, robotics and its application
|high-resolution cytometry; fluorescence in situ hybridization; interphase nuclei; fluorescence microscopy; automated microscopy; image analysis; 3-D analysis
|The recently developed technique of high-resolution cytometry (HRCM) enables automated acquisition and analysis of FISH stained cell nuclei using wide-field fluorescence microscopy. The method has now been extended to confocal imaging and offers the opportunity to combine the advantages of confocal and wide-field modes. Using the combined confocal and wide-field HRCM technique, it is possible to take advantage of both imaging modes. Images of some dyes (such as small hybridization dots or counterstain images of individual interphase nuclei) do not require confocal quality and can be acquired quickly in wide-field mode. On the contrary, images of other dyes (such as chromosome territories or counterstain images of cells in tissues) do require improved quality and are acquired in confocal mode. The dual-mode approach is 2-3 times faster compared with the single-mode confocal approach and the spectrum of its applications is much broader compared with both single-mode confocal and single-mode wide-field systems. The combination of high speed specific to the wide-field mode and high quality specific to the confocal mode gives optimal system performance.