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Updated: Aug 25, 2026
Immunostaining for DNA Modifications: Computational Analysis of Confocal Images
Published on: September 7, 2017
Multiple excitation confocal analysis of targets in nuclei of cytogenetic preparations
Edmond Kahn1, Aline Receveur, Philippe Coullin
1Institut National de la Santé et de la Recherche Médicale U494, Centre Hospitalier Universitaire Pitié-Salpêtrière, 75634 Paris, France. kahn@imed.jussieu.fr
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.
