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Preface
Second-harmonic generation (SHG) microscopy began to emerge as a high-resolution optical imaging
modality in the late 1990s, about 10 years aer two-photon excited uorescence (TPEF) microscopy
was pioneered by the laboratory of Watt Webb. Concurrent with two-photon microscopy, the initial
suggestion for SHG imaging was made by Colin Sheppard. Although largely unnoticed by the biomedi-
cal optics community, Isaac Freund and coworkers published a ber alignment study in rat-tail ten-
don using SHG polarization analysis, and while this was performed at low resolution (~50 μm), this
landmark study captured the essence of using SHG to ...