Commercially viable polymer electrolyte membrane fuel cell (PEMFC) systems for transportation and stationary power applications require optimal combination of performance and durability at a competitive cost. Optimized water management across a fuel cell is key to improving performance and maintaining high conductivity of the ionomer in PEM and catalyst layers (CLs), while preventing excessive flooding in CLs, gas diffusion layers (GDLs), channels, and manifolds. Durability is also greatly affected by water transport as certain degradation mechanisms are accelerated depending on the humidity levels. As well, catalyst layers that suffered carbon corrosion due to long operation and/or high number of starts/stops are more susceptible to flooding. Imaging in situ and ex situ enables crucial insight into the microstructure of the fuel cell components and materials, associated water transport, and their effect on cell performance, dynamic response, and degradation mechanisms. Several case studies are presented to highlight the use of optical imaging, neutron imaging, and micro X-ray computed tomography (microXCT) to investigate fuel cell components across a range of length scales. Laboratory-scale microXCT instruments are used ex situ to resolve the three-dimensional structure of the GDL and electrodes, before and after various accelerated stress tests. Optical imaging in operating fuel cells offers a cost-effective combination of high temporal resolution (tens of frames per second) and high spatial resolution (several µm). The imaging requires special cell design with visual access to the imaged area (GDL surface and channels [1] or catalyst layer surface [2]). We employed optical visualization in a variety of studies, such as to visualize liquid water dynamics in the cathode and anode flow fields, or at the interface between the CL and the GDL. Besides water transport studies, optical imaging can quantify in-plane gas distribution in the flow fields during startups and shutdowns [3]. Neutron imaging is a powerful tool to visualize and quantify water transport in operating fuel cells. It has a high sensitivity to small amounts of water inside a cell, while having high transmission through the common fuel cell hardware. At a lower spatial resolution of 250 µm, neutron imaging is used to measure in-plane water distribution with high temporal resolution and large field of view of up to 20 cm by 20 cm. We studied the performance with novel NSTF (nano structured thin film) electrodes and GDL materials [4], and also visualized water and ice formation when cell was subjected to repeated starts at sub-freezing temperatures [5]. Since neutron imaging provides water content integrated along the neutron beam, we combined neutron imaging with the optical visualization to study water transport in different flow field geometries [6] as well as in an operating electrolyzer [7]. Such approach with simultaneous imaging provides additional information about the water location, and in certain situations allows distinguishing between channel and GDL water, or between anode and cathode channels and manifolds. High-resolution neutron imaging is able to measure water distribution across the cell thickness at spatial resolution as high as 10 µm. Image processing procedure is developed to measure the water uptake and observe Schroeder’s paradox in situ in PEMs [8,9]. Further, properties of the microporous layer (MPL) of the GDL were manipulated to prevent excessive flooding in cathode catalyst layers. Anode flooding is evidenced by optical visualization, simultaneous imaging, and high-resolution neutron imaging. Increased water transport across the membrane, from cathode to anode, can be detrimental as liquid water on the anode side may cause localized fuel starvation and cause irrecoverable degradation, similar to accelerated carbon corrosion during unassisted startups and shutdowns. However, for certain novel cathodes, which are prone to flooding, e.g. nano-structured thin film (NSTF) electrodes and non-precious group metal (non-PGM) catalysts, water removal through the anode has proven to be a viable option to improve the performance by reducing cathode water content. The authors acknowledge support from US Department of Energy EERE FCTO, Los Alamos National Laboratory LDRD (Laboratory Directed Research and Development) program, Federal Transit Administration, US Department of Commerce, and NIST Center for Neutron Research.