Many two photon users have a large span of settings available to them that results from supplier implementation based on academic advancement in the field. However, optimized parameters is seldomly used du to the lack of knowledge and transmission towards non-expert users. This workshop aims to address one of those gaps by exploiting a common feature available on modern ultrafast laser allowing pre-compensation of pulse duration. We will investigate the effect of group delay dispersion (GDD) compensation on a thick sample at various depths and its impact on collected fluorescent signal, photobleaching and phototoxicity and compare with a more conventional laser power compensation. We will further investigate this method’s relevance on second harmonic generation signal.