Time reversal optical tomography (TROT) approach is used to detect and locate absorptive targets embedded in a highly scattering turbid medium to assess its potential in breast cancer detection. TROT experimental arrangement uses multi-source probing and multi-detector signal acquisition and Multiple-Signal-Classification (MUSIC) algorithm for target location retrieval. Light transport from multiple sources through the intervening medium with embedded targets to the detectors is represented by a response matrix constructed using experimental data. A TR matrix is formed by multiplying the response matrix by its transpose. The eigenvectors with leading non-zero eigenvalues of the TR matrix correspond to embedded objects. The approach was used to: (a) obtain the location and spatial resolution of an absorptive target as a function of its axial position between the source and detector planes; and (b) study variation in spatial resolution of two targets at the same axial position but different lateral positions. The target(s) were glass sphere(s) of diameter ~9 mm filled with ink (absorber) embedded in a 60 mm-thick slab of Intralipid-20% suspension in water with an absorption coefficient μa ~ 0.003 mm-1 and a transport mean free path lt ~ 1 mm at 790 nm, which emulate the average values of those parameters for human breast tissue. The spatial resolution and accuracy of target location depended on axial position, and target contrast relative to the background. Both the targets could be resolved and located even when they were only 4-mm apart. The TROT approach is fast, accurate, and has the potential to be useful in breast cancer detection and localization.© (2011) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.