|Abstract||Optical microscopes have proven their use as a powerful tool for studying biological samples. In spite of many successful applications there are still many obstacles limiting microscopy applications. Besides the resolution limitation, background and probe toxicity are two main aspects limiting many potential applications. Any new method that would allow an increase in signal-to-background ratio and at the same time reduce probe photobleaching and phototoxicity would immediately allow many new investigations like, for example, studying long lasting processes such as embryo development, cancer drug delivery, or plant growth. In our approach, we combine a technique called single plane illumination microscopy (SPIM) with multi-pulse pumping in order to enhance the signal-to-background ratio and at the same time reduce light exposure times for the specimen. To test applicability and limits of the new technology, we developed a phantom model containing various probes and an artificial background.