DBD
Cell suspension (DNA damaged)
Microgel embedding
Alkaline unwinding
Neutralization and Protein removal
D
Washing
Dehydration
D
FISH
Incubation with DNA probe(s)
Washings and Detection
Image capture and Digital image analysis
22 DNA Breakage Detection-FISH (DBD-FISH)
283
Fig. 1. Outline of the DBD-FISH
protocol
pose, e.g. in composition, conditions and order of incubation in the unwinding and/or lysing solutions.
Materials
- Standard and low melting point agarose
- Epifluorescence microscope with appropriate filters and objectives
(e.g. Leica DMRB)
- High-sensitivity CCD digital camera (e.g. Ultrapix-1600, Astrocam,
Cambridge, UK)
- Image analysis software (e.g. Visilog 5.1, Noesis, France)
- Alkaline unwinding solution (pH 12.2), freshly prepared
Solutions