Wheat (Triticum aestivum L.) breeding programs use doubled haploid (DH) production methods to shorten the time required to develop new varieties. Haploid plants are generated and the chromosomes are doubled with anti-mitotic agents to create fully homozygous doubled haploids. Since the 1990s, the maize pollen/embryo rescue technique has been used to produce wheat haploids, but it is a time consuming and labour intensive process. Instead, haploid plants can be grown from individual immature pollen cells (microspores). The Winter Wheat breeding program at the Ottawa Research and Development Centre (ORDC) evaluated a microspore culture protocol using a number of F1 crosses. Microspore culture offered greater efficiency because haploid embryos and green plants were produced from fewer spikes with microspore culture than via maize pollen/embryo rescue. Green plant formation using microspore culture was highly dependent upon the genetic background. Number of green plants produced per spike ranged from 1 to 49, and the proportion of green plants as a fraction of total plants produced varied from 9% to 83% depending on the cross used. Putative spontaneous chromosome doubling of plants ranged from 11% to 26% depending on genetic background, thus colchicine treatment is still required to obtain doubled haploids. In experiments currently underway, microspores are being cultured in media containing the anti-mitotic herbicide trifluralin in an attempt to improve the level of spontaneous chromosome doubling in vitro. With the use of flow cytometry to determine green plant ploidy prior to colchicine treatment, microspore culture has become an effective DH production tool in the AAFC Ottawa Winter Wheat breeding program.