The identification of coherent structures from experimental or numerical data is an essential task when conducting research in fluid dynamics. This typically involves the construction of an empirical mode base that appropriately captures the dominant flow structures. The most prominent candidates are the energy-ranked proper orthogonal decomposition (POD) and the frequency ranked Fourier decomposition and dynamic mode decomposition (DMD). However, these methods fail when the relevant coherent structures occur at low energies or at multiple frequencies, which is often the case. To overcome the deficit of these rigid approaches, we propose a new method termed Spectral Proper Orthogonal Decomposition (SPOD). It is based on classical POD and it can be applied to spatially and temporally resolved data. The new method involves an additional temporal constraint that enables a clear separation of phenomena that occur at multiple frequencies and energies. SPOD allows for a continuous shifting from the energetically optimal POD to the spectrally pure Fourier decomposition by changing a single parameter. In this article, SPOD is motivated from phenomenological considerations of the POD autocorrelation matrix and justified from dynamical system theory. The new method is further applied to three sets of PIV measurements of flows from very different engineering problems. We consider the flow of a swirl-stabilized combustor, the wake of an airfoil with a Gurney flap, and the flow field of the sweeping jet behind a fluidic oscillator. For these examples, the commonly used methods fail to assign the relevant coherent structures to single modes. The SPOD, however, achieves a proper separation of spatially and temporally coherent structures, which are either hidden in stochastic turbulent fluctuations or spread over a wide frequency range.