Apoptosis induced kinetic changes in autofluorescence - possible application for single cell analysis on chip

F. Wolbers, Helene Andersson, I. Vermes, Albert van den Berg

    Research output: Contribution to journalArticleAcademicpeer-review

    16 Citations (Scopus)

    Abstract

    Introduction: This paper presents a new method using natural cellular fluorescence (autofluorescence, AF) to study apoptosis. Measurement of AF reduces sample preparation time and avoids cellular toxicity due to the fact that no labelling is required. Methods: Human promyelocytic leukemic HL60 cells were incubated with camptothecin (CPT), tumour necrosis factor (TNF)-agr in combination with cycloheximide (CHX), or irradiated with 6 or 10 Gray, during varying time periods, to initiate apoptosis. AF was measured at the flow cytometer. Results: Induction of apoptosis results in the shrinkage of the cell and the fragmentation into apoptotic bodies. With flow cytometry, 4 subpopulations, viable, early apoptotic, late apoptotic and the necrotic cells, can be distinguished. Induction of apoptosis results in a decrease in AF intensity compared to untreated HL60 cells, especially seen in the late apoptotic subpopulation. The AF intensity is found to decrease significantly in time (between 2 h and 24 h) for all the four apoptotic inducers used. Conclusions: Our results show that it is possible to specifically measure the apoptotic-induced kinetic changes in AF in HL60 cells. A decrease in AF intensity is seen from 2 h till 24 h. These results open a door for future developments in single-cell analysis.
    Original languageEnglish
    Pages (from-to)749-755
    Number of pages7
    JournalApoptosis
    Volume9
    Issue number6
    DOIs
    Publication statusPublished - 2004

    Keywords

    • METIS-219071
    • IR-47985

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