Laser scanning cytometry allows detection of cell death with morphological features of apoptosis in cells stained with PI.
Laser scanning cytometry (LSC), a newly developed technology, allowed simple detection of dead cells with morphological features of apoptosis for cells stained with only propidium iodide (PI). HeLa cells were treated with Adriamycin (ADM, 0.5 or 1.0 microgram/ml). A PI fluorescence value (representing DNA content) versus PI fluorescence peak (representing chromatin condensation) cytogram of LSC made it possible to segregate cells with high PI fluorescence peak from others in a cell population and concomitantly to analyze the relationship between the cells and the cell cycle. A fraction of the cells manifesting hypercondensation of chromatin was exclusively present in a cell population treated with ADM. Visual inspection of the cells defined in the cytogram revealed morphological features of apoptosis. LSC analysis facilitates monitoring effects of anticancer drugs on a cell population, because nuclear DNA staining with PI is simple and rapid.
Furuya
T
T
Department of Pathology, Yamaguchi University School of Medicine, Ube, Japan.
Kamada
T
T
Murakami
T
T
Kurose
A
A
Sasaki
K
K
D000903:Antibiotics, Antineoplastic / Q000008:administration & dosage / Q000494:pharmacology; D017209:Apoptosis; D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology; D016923:Cell Death / Q000502:physiology*; D002467:Cell Nucleus / Q000737:chemistry / Q000187:drug effects; D004396:Coloring Agents; D004247:DNA / Q000032:analysis / Q000187:drug effects; D004317:Doxorubicin / Q000008:administration & dosage / Q000494:pharmacology; D005434:Flow Cytometry; D005453:Fluorescence; D016193:G1 Phase / Q000187:drug effects / Q000502:physiology; D006367:HeLa Cells / Q000737:chemistry*; D006801:Humans; D019044:Image Cytometry / Q000379:methods*; D007834:Lasers; D011419:Propidium; D013194:Staining and Labeling
D000903:Antibiotics, Antineoplastic; D004396:Coloring Agents; D011419:Propidium; D004317:Doxorubicin; D004247:DNA