Document Detail


Machine vision based stochastic analysis of cancer cell mitochondrial dysfunction induced by a BH3 domain.
MedLine Citation:
PMID:  18814032     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We have developed a versatile and rapid method for the quantitative estimation of cell death kinetics, following direct single-shot activation of the mitochondrial death pathway by a cell permeable BH3 activator peptide (D-R(8)BH3(BID)). This approach employs timelapse epifluorescent imaging of live cells and a machine- vision based feature extraction algorithm, to measure unidirectional stochastic transitions associated with mitochondrial inner membrane potential depolarization and/or permeability transition, at single cell resolution. This data is transformed to enable construction of a right step-wise survival function using the product limit estimator, and estimation of a median latency parameter (lambda), defined for the entire imaged cell population. Estimates of lambda computed for cells exhibiting two-colour fluorescence can be compared statistically using the Mantel-Hansel test. This general method has been applied to measure the kinetics and temporal ordering of BH3 domain induced mitochondrial depolarization and inner membrane permeabilization in cancer cells, and demonstrates the robustness of this technique in resolving temporally distinct intracellular events within individual cells.
Authors:
Alex D Chacko; Nyree T Crawford; Patrick G Johnston; Dean A Fennell
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Apoptosis : an international journal on programmed cell death     Volume:  13     ISSN:  1573-675X     ISO Abbreviation:  Apoptosis     Publication Date:  2008 Nov 
Date Detail:
Created Date:  2008-10-14     Completed Date:  2008-12-03     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9712129     Medline TA:  Apoptosis     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1386-93     Citation Subset:  IM    
Affiliation:
Centre for Cancer Research and Cell Biology, Queen's University Belfast, 97 Lisburn Road, Belfast BT97BL, UK.
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MeSH Terms
Descriptor/Qualifier:
Algorithms
Apoptosis
Biophysics / methods
Circular Dichroism
Cytochromes c / metabolism
Humans
Kinetics
Membrane Potentials
Microscopy, Fluorescence / methods
Mitochondria / metabolism*
Models, Biological
Protein Conformation
Protein Structure, Tertiary
Stochastic Processes
Time Factors
Chemical
Reg. No./Substance:
9007-43-6/Cytochromes c

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


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