This paper describes the adaptation of the laser-induced fluorescence measurement technique for the investigation of the primary breakup of modern diesel and gasoline direct injection sprays. To investigate the primary breakup, a microscopic technique is required, and with the help of special tracer dyes, a high fluorescence signal can be achieved in the visible range of the electromagnetic spectrum, resulting in good image quality with a nonintensified camera. Besides the optimization of the optical setup for the microscopic field of view, different tracer dyes are compared, and their solubility and fluorescence are tested in the desired surrogate and real-world fuels. As a tracer, the phenoxazine dye Nile Red was found to provide sufficient solubility in alkanes as well as suitable emission and excitation spectrum for the use of the second-harmonic frequency of a Nd:YAG laser (532 nm). The good quantum efficiency delivered by Nile Red also meant that single-shot images clearly showing spray structures in regions measuring up to 3 mm by 3 mm around the nozzle outlet could be recorded. Compared to relatively easy shadowgraph techniques and complex and costly x-ray synchrotron measurements, light sheet fluorescence microscopic imaging is not overly complex yet delivers excellent data on spray structures as well as qualitative fuel distribution.