Videomicrofluorometry on living cells and discriminant factorial analysis to study cell cycle distributions.
After a rapid overview of the approaches used to study cell cycle, a fluorescent digital imaging microscopy method is proposed. This method is improved by a factorial analysis relying on the evaluation of several parameters recorded on each living cell. Single lympho-blastoid living cells are labeled with three fluorescent markers: Hoechst 33342 for nuclear DNA, Rhodamine 123 for mitochondria and Nile Red for plasma membrane. For each cell, morphological and functional information parameters are obtained. A typological analysis is used to separate control cells into four groups: G0-G1, S, G2+M and polyploid cells Gn. These control cells define a learning population used to analyze untreated and adriamycine treated cells as supplementary individuals in a discriminant factorial analysis. Such an approach allows to accurately evidence the change of the values of some cellular parameters.
Savatier
J
J
Biophysique et Dynamique des Systèmes Intégrés, Université de Perpignan, Perpignan Cedex, France.
Gbankoto
A
A
Vigo
J
J
Salmon
J M
JM
D001562:Benzimidazoles / Q000737:chemistry; D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D003592:Cytophotometry / Q000379:methods; D004317:Doxorubicin / Q000494:pharmacology; D005163:Factor Analysis, Statistical; D006801:Humans; D007091:Image Processing, Computer-Assisted; D047569:Laser Scanning Cytometry / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods; D018715:Microscopy, Video / Q000379:methods; D010078:Oxazines / Q000737:chemistry; D020112:Rhodamine 123 / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods
D001562:Benzimidazoles; D010078:Oxazines; D020112:Rhodamine 123; D004317:Doxorubicin; C044808:nile red; C017807:bisbenzimide ethoxide trihydrochloride