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Qualitative and quantitative analysis of DNA fragmentation using digital imaging
Y Vodovotz1, A Hsing, J A Cook
1Radiation Biology Branch, National Cancer Institute, Bethesda, Maryland 20892, USA. yoram@box-y.nih.gov
Analytical Biochemistry
|August 1, 1997
Summary
Quantifying apoptosis, a common cell death pathway, is challenging. This study introduces RIT120 software for rapid, objective analysis of DNA fragmentation, distinguishing apoptotic cell death from necrosis.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Apoptosis is a critical cellular process, but its accurate quantification, especially alongside necrosis, presents analytical difficulties.
- Distinguishing between apoptosis and necrosis is essential for understanding cellular fate and disease progression.
Purpose of the Study:
- To present a novel method for the quantitative and qualitative analysis of apoptosis.
- To demonstrate the utility of RIT120 digital imaging software for analyzing apoptotic DNA ladders.
Main Methods:
- Utilized RIT120 software for densitometric scanning of DNA ladder autoradiographs.
- Employed quantitative subtraction of necrotic DNA degradation from apoptotic DNA fragmentation.
- Integrated peak areas from densitometric curves to quantify apoptosis.
Main Results:
- Successfully quantified apoptosis induced by various agents including serum, TNF-α, TGF-β1, and nitric oxide.
- The RIT120 software enabled objective measurement of DNA fragmentation patterns.
- The method allowed for the differentiation of apoptotic DNA ladders from general DNA degradation.
Conclusions:
- RIT120 software offers a rapid, objective, and cost-effective solution for quantifying apoptosis.
- This approach simplifies the analysis of apoptotic DNA ladders, even in the presence of necrosis.
- The described method provides a valuable tool for researchers studying cellular death pathways.