Multiple excitation confocal analysis of targets in nuclei of cytogenetic preparations.

OBJECTIVE

To demonstrate that cellular preparations requiring color analysis of different domains stained by molecular cytogenetic methods (fluorescence in situ hybridization) can be processed by spectral analysis of fluorescent emissions by either factor analysis of medical image sequences (FAMIS) or a META confocal configuration to isolate fluorescent probes.

STUDY DESIGN

Three-dimensional sequences of images obtained by spectral analysis in a META confocal microscope (Carl Zeiss SAS, Jena, Germany) were analyzed by META processing and the FAMIS algorithm, which provides factor curves. META and factor images were then the result of image-processing methods that cover emission spectra.

RESULTS

Factor curves and factor or META images can help to analyze targets inside nuclei.

CONCLUSION

It is possible to process preparations containing numerous spots on different colors to differentiate stained targets and to improve visualization and detection.