The unconformity-related uranium (URU) deposits in the Proterozoic Athabasca Basin are one of the most important U resources in the world. This type of U deposit can be divided into monometallic (U) and polymetallic (U-Ni-Co-As) subtypes. While it is generally agreed that the URU deposits formed from reaction between oxidizing, basinal brines carrying U and/or Ni-Co-As with reducing basement fluids or lithologies, it is debatable whether the polymetallic deposits formed from co-enrichment of U-Ni-Co-As or enrichment of U superimposed by a separate Ni-Co-As mineralization event. This study addresses this problem through mineralogical, geochemical and fluid inclusion investigation of the Midwest U-Ni-Co-As deposit. Petrographic studies indicate that the sequence of ore precipitation started with uraninite, followed by Ni-Co arsenides and sulfoarsenides and then Cu-Pb-Fe sulfides, and this sequence was repeated episodically. This observation suggests that the deposit did not form from two separate U and Ni-Co-As mineralization events, but rather multiple episodes of U-Ni-Co-As mineralization. Linear correlations between chemical ages and Si-Ca-Fe contents of the most pristine uraninite U1 suggest a maximum primary mineralization of ca. 1600 Ma, which is consistent with the inferred primary U mineralization age in the Athabasca Basin. Microthermometric and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) analyses of fluid inclusions in syn-mineralization drusy quartz indicate that the composition of the ore-forming fluids is characterized by the H2O-NaCl-CaCl2-KCl-MgCl2 system and comparable to those from both monometallic and polymetallic URU deposits. The relationship between U and Ni + Co in the fluid inclusions and its comparison with other URU deposits support a model in which U and Ni-Co were co-enriched in a unified mineralization process. The development of breccia structures in the ores and the dramatic fluid pressure fluctuation revealed by fluid inclusions suggest that the deposit formed from multiple episodes of fluid flow related to repeated reactivation of basement-rooted faults.