Notch is actively involved in various life processes including osteogenesis. The role of Notch signalling in the terminal mineralisation is largely unknown. It was noted that Hes1, a downstream target of Notch signalling was highly expressed in mature osteocytes compared to osteoblasts. Using a recently developed thermolabile cell line (IDG-SW3) and Rbpj luciferase reporter, we demonstrated that DMP-1 expression was inhibited and mineralization process was significantly altered when Notch pathway inactivated by addition of DAPT. Dysregulation of notch in osteocyte differentiation can result in spontaneous deposition of calcium phosphate on collagen fibrils, disturbed transportation of intracellular mineral vesicles, altered crystal structure of minerals observed by SAED, decreased bonding force between minerals and substrates tested by AFM, and dendrite development inhibition with decreased expression of E11.  In conclusion, the evidence presented here suggests that Notch plays a critical role in osteocyte differentiation and biomineralisation.