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While research on racial segregation in cities has grown rapidly over the last several decades, its foundation remains the analysis of the neighbourhoods where people reside. However, contact between racial groups depends not merely on where people live, but also on where they travel over the course of everyday activities. To capture this reality, we propose a new measure of racial segregation – the segregated mobility index (SMI) – that captures the extent to which neighbourhoods of given racial compositions are connected to other types of neighbourhoods in equal measure. Based on hundreds of millions of geotagged tweets sent by over 375,000 Twitter users in the 50 largest US cities, we show that the SMI captures a distinct element of racial segregation, one that is related to, but not solely a function of, residential segregation. A city’s racial composition also matters; minority group threat, especially in cities with large Black populations and a troubled legacy of racial conflict, appears to depress movement across neighbourhoods in ways that produce previously undocumented forms of racial segregation. Our index, which could be constructed using other data sources, expands the possibilities for studying dynamic forms of racial segregation including their effects and shifts over time.