Let slides dry 20 minutes to 3 hours. Sterile filter and ice. Denature probes by heating to 80 degrees C for 5 minutes. Chill on ice. Pipet carefully onto tissue and coverslip with Hybrislip plastic cover slips.
Place slides horizontally in slide mailers.
Do not mix slides containing different probes in the same slide mailer, because cross-contamination may occur. Hybridize in horizontal slide mailers in a humidified box at 65 degrees C for 16 to 24 hours. Develop at room temperature in the dark for 10 minutes to 3 days, depending on abundance of transcript. Substitution of FITC-labeled secondary antibodies is not recommended. Only Alexa488 has worked in my hands.