A simple theoretical framework to compute the eigenvalues of a cylindrically symmetric prolate diffusion tensor (D) from one of the rotationally-invariant diffusion anisotropy indices and average diffusivity is presented and validated. Cylindrical or axial symmetry assumes a prolate ellipsoid shape ( ሻ ؍ 1 > Ќ ؍ ( 2 ؉ 3 )/2; 2 ؍ 3 ). A prolate ellipsoid with such symmetry is largely satisfied in a number of white matter (WM) structures, such as the spinal cord, corpus callosum, internal capsule, and corticospinal tract. The theoretical model presented is validated using in vivo DTI measurements of rat spinal cord and human brain, where eigenvalues were calculated from both the set of diffusion coefficients and a tensor analysis. This method was used to retrospectively analyze literature data that reported tensor-derived average diffusivity, anisotropy, and eigenvalues, and similar eigenvalue measurements were obtained. The method provides a means to retrospectively reanalyze literature data that do not report eigenvalues. Diffusion tensor imaging (DTI) is a sensitive, noninvasive method that is increasingly being used to follow subtle changes in tissue microstructural organization (1-5) due to development (6 -8), cortical maturation (9 -11), normal aging (12-15), traumatic and diffuse axonal injuries (16), neurodegenerative and neurovascular diseases (17-19), drug abuse (20), and therapeutic intervention (21). DTI provides important qualitative and rotationally-invariant quantitative information about the integrity of compact fiber bundles, and may provide more sensitive and specific surrogate markers of disease activity, therapy, and clinical outcomes (1,6,20 -22). DT anisotropy is expressed as a single scalar index, such as fractional anisotropy (FA) (23) or relative anisotropy (RA), which is also known as A (24). A has also been termed normalized relative anisotropy (RA n ϭ 1/͌2RA ϭ A ) (25). While reduced diffusion anisotropy is interpreted as an indication of compromised fiber integrity, its specificity is somewhat limited (3-5).For instance, reduced anisotropy can result from excessive partial volume averaging from differently oriented fibers (1-5), edema (12), cerebrospinal fluid (CSF) (13), compromised myelin structure, changes in axonal morphology, or altered interaxonal spacing of fiber bundles (4,6,(15)(16)(17)(18)(19)(20)(21).Normal tissue maturation and development (6,9 -11), aging (12-15), pathology, injury, and recovery (16 -28) involve specific directional changes to the axonal structure and myelination (1)(2)(3)(4)(5)15,22,25). Indeed, recent studies have accumulated considerable evidence that individual DT eigenvalues provide enhanced pathological specificity compared to anisotropy and mean diffusivity indices (23-29). Beaulieu (4) summarized a number of studies prior to 2002 that highlighted the pathological specificity of the individual diffusivities. More recent DTI investigations using animal models of human diseases concluded that the tensor eigenvalues are more specific markers of m...