The aim of this study was to develop an effective method of overwintering reserve honey bee queens in two-storey mini-plus mating nuclei and in 3-comb nuclei (frames 36 x 26 cm, Wielkopolski hive). The assay was performed during three wintering seasons (2005 - 2008) parallel at two centers in Poland: the Division of Apiculture at the University of Life Sciences (SGGW) in Warsaw, and the Apiculture Division at the University of Warmia and Mazury (UWM) in Olsztyn. The results showed that 59% of queens overwintered in mini-plus nuclei and 77% in 3-comb nuclei. Among queens in mini-plus nuclei 63% overwintered in bee yard and only 55% in cellar. Within queens in 3-comb nuclei, 62% overwintered in Olsztyn and 91% in Warsaw. The highest survival rate of 93% was observed in Warsaw during the first season. Due to low survival rate, it is not recommended to overwinter the queens in miniplus nuclei.