Recently, structural assessments, particularly of the TM, have occurred in a more native three-dimensional context with two-photon multimodal imaging. Two photon-microscopy (TPM) enabled high-resolution deep tissue optical sectioning in whole fixed or live tissue . Different modalities, such as autofluoresence and second harmonic generation demonstrated native trabecular anatomy and allowed for segregation of different extracellular matrix components such as collagen and elastin [31–33]. This type of tissue-based trabecular analyses has led to exploration of live cell dynamics and study of drug effects for IOP modulation. TPM of AHO anatomy, however, is currently not applied in living human individuals.