Roses are known for their low pollen quality. It remains unclear how many days of pollen should be used in order not to reduce crossbreeding success. The study was conducted to determine whether short-term pollen storage is effective on the crossing success of roses comparatively in vivo and in vitro. The study was conducted in a greenhouse and a cytology laboratory at Ankara University, Turkey. The 'Avalanche' rose variety was used as the seed parent, and 'Magnum' was the pollen parent. A total of 210 crosses were made. Pollens were used immediately (day 0) and stored at +4°C for 6 days after collection. The pollen viability by IKI and the germination rates by the petri dish method, the fruit and seed set, the fruit and seed weight, and the seed germination rate were recorded. All examined traits showed a decreasing trend as the storage time increases. The pollen germination rates were recorded at 1.63% for stored pollen at six days and 15.27% for fresh pollen. The highest fruit and seed set rates were obtained from crossing with pollen stored for one day. The fruit set decreased below 10% as of the 5th day, and the seed numbers decreased 1.8 times compared to one day of storage. No germination was observed in seeds obtained from crosses with pollen stored for 5 or 6 days. A positive correlation was found between seed number and pollen germination rate. The study concluded that successful crossing could be made with pollen stored at +4°C for up to 3 days.