This dataset contains breakthrough curves of conservative (fluorescein) and reactive (resazurin and resorufin) tracers resulting from instantaneous tracer experiments in a lowland agricultural stream. Breakthrough curves were measured seasonally at four locations within the stream, creating three experimental reaches, in the Wood Brook, Staffordshire from July 2016 to March 2017. Breakthrough curves were measured in-situ using on-line fluorometers configured to measure the excitation of fluorescein, resazurin and resorufin every 10 seconds. The breakthrough curves were measured to determine hydrological metrics of advective transport, transient storage and aerobic respiration. The work was funded by the Natural Environment Research Council, UK through a through a Central England NERC Training Alliance Studentship and grant NE/L004437/1, with additional funding provided by the European Union through the H2020-MSCA-RISE-2016 project 734317.