In Situ Detection of DNA Strand Breaks in Analysis of Apoptosis by Flow- and Laser-Scanning Cytometry
The presence of a multitude of DNA strand breaks resulting from fragmentation of nuclear DNA by the caspase-activated DNase is one of the most chaacteristic features of apoptotic cells (1 ,2 ). A widely used methodology to detect apoptotic cells, thus, relies on labeling DNA strand breaks in situ, within the nuclear chromatin, either with fluorochromes (3 –5 ) or absorption dyes (6 –8 ). The overview of the techniques (TUNEL techniques), which were developed independently by Gavrieli et al., (6 ) and by us (3 –5 ) is presented in Chapter 3 . One advantage of strand break labeling with fluorochromes is that such cells can rapidly be analyzed by flow cytometry. When cellular DNA content also is measured in these cells, the bivariate analysis of such data provides information about DNA ploidy or the cell cycle phase specificity of apoptosis (3 ,9 ).
- DNA-Unwinding Test Using Eukaryotic DNA Topoisomerase I
- Isolation of Bacterial Cell Membranes Proteins Using Carbonate Extraction
- Quantifying the Titer and Quality of Adenovirus Stocks
- Generating Mammalian Sirtuin Tools for Protein-Interaction Analysis
- Probe Ordering and Distancing by FISH
- In-Frame Cloning of Synthetic Genes Using PCR Inserts
- Gene Expression Profiling in Formalin-Fixed, Paraffin-Embedded Tissues Using the Whole-Genome DASL Assay
- In Vitro Transcription/Translation Analysis for the Identification of Translation-Terminating Mutations
- Cationic Liposome-Mediated DNA Delivery to the Lung Endothelium
- Construction of a Library of Random Mutants in the Spirochete Leptospira Biflexa Using a Mariner Transposon