The valve regulated lead acid (VRLA) battery is a predominant electrochemical storage system that stores energy in a cheap, reliable and recyclable manner for innumerable applications. The absorptive glass mat (AGM) separator is a key component, which is pivotal for the successful functioning of the VRLA battery. Herein, the intricate three-dimensional (3D) porous structure of AGM separators has been unveiled using X-ray micro-computed tomography (microCT) analysis. X-ray microCT has quantified a variety of fiber and structural parameters including fiber orientation, porosity, tortuosity, pore size distribution, pore interconnectivity and pore volume distribution. A predictive model of hydraulic tortuosity has been developed based upon some of these fiber and structural parameters. Moreover, the pore size distribution extracted via X-ray microCT analysis has served as a benchmark for making a comparison with the existing analytical model of the pore size distribution of AGM separators. Pore size distributions obtained via the existing analytical model and through X-ray microCT analysis are in close agreement.