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.


METHODS
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.


CONCLUSIONS
It is possible to process preparations containing numerous spots on different colors to differentiate stained targets and to improve visualization and detection.
Kahn
Edmond
E
Institut National de la Santé et de la Recherche Médicale U494, Centre Hospitalier Universitaire Pitié-Salpêtrière, 75634 Paris, France. kahn@imed.jussieu.fr
Receveur
Aline
A
Coullin
Philippe
P
Frouin
Frédérique
F
Blanco-Soulas
Catherine
C
Todd-Pokropek
Andrew
A
Bernheim
Alain
A
D002292:Carcinoma, Renal Cell / Q000235:genetics / Q000473:pathology; D002467:Cell Nucleus / Q000737:chemistry / Q000235:genetics*; D002879:Chromosomes, Human, Pair 10 / Q000737:chemistry / Q000235:genetics; D002880:Chromosomes, Human, Pair 11 / Q000737:chemistry / Q000235:genetics; D002895:Chromosomes, Human, Pair 5 / Q000737:chemistry / Q000235:genetics; D002896:Chromosomes, Human, Pair 6 / Q000737:chemistry / Q000235:genetics; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008214:Lymphocytes / Q000737:chemistry / Q000187:drug effects; D008297:Male; D018613:Microscopy, Confocal / Q000379:methods; D010835:Phytohemagglutinins / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D005456:Fluorescent Dyes; D010835:Phytohemagglutinins