IQ-FISH analysis was performed using an IQ-FISH Fast Hybridization Buffer, Sure FISH PML-RARA Dual fusion probe (Agilent Technologies , West Cedar Creek, TX) and a Dako Cytology FISH Accessory Kit (Dako, Glostrup, Denmark) containing Pretreatment buffer, Wash buffer and Stringent wash buffer. The sample slides were chemically aged by treatment with pretreatment buffer at 95°C for 10 min and then cooled at room temperature for 10 min.
The slides were dehydrated through an ethanol series (70%, 85%, and 100%), and the probe mixture was then applied. The slides were sealed with coverslips, denatured at 66°C for 10min, and incubated at 45°C for 1 h. Thereafter, the slides were washed with Wash buffer and Stringent wash buffer according to the manufacturer's instructions. The nuclei were stained and mounted with DAPI II counterstain (Abbott Molecular/Vysis. Des Plaines, IL) and VECTASHIELD Mounting Medium ( Vector Laboratories, Burlingame, CA) . The signals obtained by IQ FISH were analyzed as described for conventional FISH.
The slides were dehydrated through an ethanol series (70%, 85%, and 100%), and the probe mixture was then applied. The slides were sealed with coverslips, denatured at 66°C for 10min, and incubated at 45°C for 1 h. Thereafter, the slides were washed with Wash buffer and Stringent wash buffer according to the manufacturer's instructions. The nuclei were stained and mounted with DAPI II counterstain (Abbott Molecular/Vysis. Des Plaines, IL) and VECTASHIELD Mounting Medium ( Vector Laboratories, Burlingame, CA) . The signals obtained by IQ FISH were analyzed as described for conventional FISH.