Document Detail


Apoptosis induced kinetic changes in autofluorescence of cultured HL60 cells-possible application for single cell analysis on chip.
MedLine Citation:
PMID:  15505417     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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)-alpha 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.
Authors:
F Wolbers; H Andersson; A van den Berg; I Vermes
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Publication Detail:
Type:  Comparative Study; Journal Article    
Journal Detail:
Title:  Apoptosis : an international journal on programmed cell death     Volume:  9     ISSN:  1360-8185     ISO Abbreviation:  Apoptosis     Publication Date:  2004 Nov 
Date Detail:
Created Date:  2004-10-26     Completed Date:  2005-04-28     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9712129     Medline TA:  Apoptosis     Country:  United States    
Other Details:
Languages:  eng     Pagination:  749-55     Citation Subset:  IM    
Affiliation:
Department of Clinical Chemistry, Medisch Spectrum Twente, Hospital Group, PO Box 50000, 7500 KA Enschede, The Netherlands.
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MeSH Terms
Descriptor/Qualifier:
Apoptosis / drug effects,  physiology*,  radiation effects
Camptothecin / pharmacology
Cycloheximide / pharmacology
Enzyme Inhibitors / pharmacology
Feasibility Studies
Flow Cytometry
Fluorescence*
HL-60 Cells
Humans
Kinetics
Oligonucleotide Array Sequence Analysis*
Protein Synthesis Inhibitors / pharmacology
Tumor Necrosis Factor-alpha / pharmacology
X-Rays
Chemical
Reg. No./Substance:
0/Enzyme Inhibitors; 0/Protein Synthesis Inhibitors; 0/Tumor Necrosis Factor-alpha; 66-81-9/Cycloheximide; 7689-03-4/Camptothecin

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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