X‐Ray diffraction microstrain characterization is a technique which enables the quantification of threading dislocations by measuring the radial microstrain field surrounding these defects. This work demonstrates how to accurately perform these measurements on a broad range of GaN samples. In particular, for GaN grown on Si substrates with large stress gradient through the layer, the measurements need to be performed on the (20–25) reflection to avoid being affected by a macrostrain influence. A comparison of this characterization with cathodoluminescence and X‐ray diffraction lattice misorientations measurements shows differences in the measurement depth, so the results are affected by the various evolutions of dislocations densities through the layers of the different samples. Independently of the measurement depth, the analysis also highlights a dependence of the X‐ray measurements results on the type of sample, in particular the starting substrate. Microstrain measurements appear therefore as being well suited for the comparison of threading dislocations densities between similar GaN layers. It can be used to replace lattice misorientations measurements, or be associated with them to obtain an edge versus screw dislocations ratio. This work presents an X‐ray diffraction measurement of threading dislocations densities, based on the surrounding microstrain field analysis. It demonstrates how to accurately characterize different gallium nitride samples, including specimens grown on silicon substrates with large stress gradient through the layer. A comparison between microstrain, lattice misorientations, and cathodoluminescence measurements highlights the potential and the issues of this technique.