Fluorescence in situ hybridization (FISH) permits the detection of unique DNA sequences of target DNA within single cells. FISH allows the identification of DNA sequences within individual cells and readily identifies genetic changes within a mixed population of cells. However, problems arise in maintaining nuclear morphology, particularly following harsh denaturation procedures where cells are heated to 80°C, causing the nuclear boundary to become blurred or the cell to completely burst. In an attempt to preserve cellular morphology and high hybridization efficiency, particularly in fresh tissue samples, we evaluated a FISH protocol using alkaline denaturation in place of heat.
Original Publication Citation
Aridgides, L. J., Stacey, M., Brihn, L., Scott, D., & Osgood, C. (2002). Fluorescence in situ hybridization on sperm using alkaline denaturation. Biotechniques, 33(2), 266-267.
Aridgides, L. J.; Stacey, M.; Brihn, L.; Scott, D.; and Osgood, C., "Fluorescence In Situ Hybridization on Sperm Using Alkaline Denaturation" (2002). Bioelectrics Publications. 211.