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.
Biophysique et Dynamique des Systèmes Intégrés, Université de Perpignan, Perpignan Cedex, France.
J
J
Savatier
A
A
Gbankoto
J
J
Vigo
J M
JM
Salmon
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