Because fatigue crack initiation and propagation of microstructurally short cracks represent most of the fatigue life in high-strength materials, especially under low amplitude loading, the study of crack initiation is of significant importance and attracted increasing attention recently. As long as the microcrack size is comparable with the grain size, the microcrack initiation is strongly influenced by the crystallographic microstructure. The main purpose of this work is to study the influence of the microstructure and variable amplitude loading on cyclic plasticity and microcrack initiation in stainless steel 304L. The first part of this work aims at enriching the crystal plasticity code Cristal ECP to better simulate the evolution of dislocation densities on slip systems and formation of PSBs which are responsible for the fatigue cyclic hardening/softening behavior. The second part concerns the evaluation of the influence of local values of mechanical factors, issued from both macroscopic and microscopic fatigue criteria, on crack initiation in the grains through the comparison between experimental surface observations and numerical simulations of 3D aggregates with realistic crystallographic orientations. It is shown that the crystal plasticity simulation can give useful contributions to predict the crack initiation sites and severity, and help to select fatigue crack initiation criterion based on mechanical parameters which actually control the crack initiation. The last part studies the effects of the variable amplitude loading (high-low sequence) on cyclic plasticity and crack initiation. Both load- and strain-controlled fatigue tests were considered. Load-controlled tests present much longer lives (6 to 9 times) than in strain-controlled tests due to strain hardening. Furthermore, simulations show that under strain-controlled loading, memory effect of overload is dependent on the amplitude level of the low amplitude block, which is more significant in high cycle fatigue than in low cycle fatigue.