Fuel cells are a promising energy conversion technology compatible with developing renewably sourced primary energy distribution. Proton exchange membrane (PEM) fuel cells are particularly suitable for automotive and portable applications. The present thesis advances novel PEM fuel cell porous transport layer (PTL) characterization and materials research. These layers link macro and nano scales by mediating energy and mass transport between reactant distribution channels and catalyst layers. Contemporary commercial PTLs are limited in selection. Moreover, typical characterization methods ignore essential material anisotropy. Herein, a novel transport layer synthesis concept is introduced. By adapting electrospinning technology, structures with engineered morphology are created. PTLs are produced with fibre diameters from 0.2 to 1.6 µm, and are characterized experimentally ex-situ and in-situ. Electrospun PTLs are shown to deliver 85% of equivalent commercial PTL current densities. Furthermore, the state-of-the-art for electronic resistance measurement of PTLs is improved, with rigorous attention given to the anisotropy of the fibre-based media. Novel method and apparatus provide this information as a function of mechanical strain. PTL in-plane resistivities are a unique contribution, where for commercial materials 4.5x10⁻⁴ to 1.5x10⁻⁴ Ω∙m are observed for strains from 0.0 to -0.5 m∙m⁻¹. Finally, electrospun PTLs are developed to investigate the effect of within-plane anisotropy upon fuel cell performance. Electrospun layers are produced with progressively greater fibre alignment to effect anisotropy. This anisotropy is visualized via microscopy, and quantified using the aforementioned electronic resistivity methods. In-situ results with electrospun PTLs, of anisotropy ratios from 1 to 6, suggest greater performance with average fibre alignment perpendicular to gas distribution channels. The present thesis’ contributions strengthen development of a PTL structure-propertyperformance relationship. With integration into a cell-level relationship, this can empower rational PEM fuel cell design