Towards multimodal nonlinear optical tomography – experimental methodology

All-optical microspectroscopic and tomographic tools reveal great potential for clinical dermatologic diganostics, i.e., investigation of human skin and skin diseases. While optical-coherence tomography has been complemented by two-photon fluorescence tomography and second-harmonic generation tomography, a joint study of various nonlinear optical microspectroscopies, i.e., application of the recently developed multimodal approach, to sizable human-tissue samples has not been evaluated up to now. Here, we present such multimodal approach combining different nonlinear optical contrast mechanisms for imaging, namely two-photon excited fluorescence (TPF), second-harmonic generation (SHG) and coherent anti-Stokes Raman scattering (CARS) into a joint microscopic experiment. We show the potential of imaging large skin areas and discuss the information obtained in a case study comparing normal skin and keloid tissue.