Despite the very simple geometry, T-shaped micro-mixers are characterized by different and complex laminar flow regimes. In the present work, experiments and direct numerical simulations are employed jointly to investigate unsteady periodic flow regimes, viz. the asymmetric and symmetric regimes. The first is characterized by a periodic dynamics of the three-dimensional structures and by a high degree of mixing, while in the second the flow always maintains a double mirror symmetry in the mixing channel, which causes a dramatic decrease of the mixing performance. A methodology, allowing us to quantitatively compare the numerical predictions with experimental flow visualizations, is used to investigate these unsteady regimes and to evaluate the relevant frequencies and the degree of mixing. In both regimes the characteristic non-dimensional frequency, based on the bulk velocity and hydraulic diameter of the mixing channel, increases with the Reynolds number, but a significant discontinuity is found at the transition from the first to the second regime.

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