Polydisperse fiber networks are the basis of many natural and manufactured structures, ranging from high-performance biobased materials to components of living cells and tissues. The formation and behavior of such networks are given by fiber properties such as length and stiffness as well as the number density and fiber-fiber interactions. Studies of fiber network behavior, such as connectivity or rigidity thresholds, typically assume monodisperse fiber lengths and isotropic fiber orientation distributions, specifically for nano scale fibers, where the methods providing time-resolved measurements are limited. Using birefringence measurements in a microfluidic flow-focusing channel combined with a flow stop procedure, we here propose a methodology allowing investigations of length-dependent rotational dynamics of nanoscale polydisperse fiber suspensions, including the effects of initial nonisotropic orientation distributions. Transition from rotational mobility to rigidity at entanglement thresholds is specifically addressed for a number of nanocellulose suspensions, which are used as model nanofiber systems. The results show that the proposed method allows the characterization of the subtle interplay between Brownian diffusion and nanoparticle alignment on network dynamics.