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This review discusses complementary nonlinear optical imaging modalities such as two\u2010photon fluorescence (TPF), second\u2010harmonic generation (SHG), and fluorescence lifetime imaging (FLIM) microscopy that together provide structural, molecular, and metabolic information. The review begins by introducing the basic principles, instrumentation, and capabilities of two\u2010photon fluorescence microscopy and FLIM. Followed by the development of SHG microscopy, highlighting advances from conventional SHG imaging to polarization\u2010resolved, circular dichroism, and Stokes\u2013Mueller polarimetric approaches. The biomedical applications of these complementary techniques are subsequently discussed, followed by an overview of multimodal imaging strategies that combine structural, molecular, and functional information. 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