Elimination of imaging artifacts in second harmonic generation microscopy using interferometry

Interferometric second harmonic generation microscopy.

We describe a novel second harmonic generation (SHG) microscope that employs heterodyne detection by interfering the epi directed SHG from a sample with SHG from a reference crystal. In addition, the microscope provides complementary reflectance information based on optical coherence microscopy (OCM). The instrument features dual balanced detection to minimize the effect of source fluctuations,...

Collinear phase matching for second harmonic generation using conoscopic interferometry

Second-harmonic imaging microscopy of living cells.

Second harmonic generation (SHG) has been developed in our laboratories as a high-resolution nonlinear optical imaging microscopy for cellular membranes and intact tissues. SHG shares many of the advantageous features for microscopy of another more established nonlinear optical technique: two-photon excited fluorescence (TPEF). Both are capable of optical sectioning to produce three-dimensional...

Two Photon Microscopy and Second Harmonic Generation

Nonlinear microscopy has become an important tool for imaging biological samples such as neurons. In two photon fluorescence, a fluorophore absorbs two photons at a low energy state, one of its electrons briefly converts to a higher energy state, and it emits a single excited photon at an energy level higher than that of the absorbed photons. In second harmonic generation (SHG), laser light is ...

Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy.

Conventionally, liver fibrosis is diagnosed using histopathological techniques. The traditional method is time-consuming in that the specimen preparation procedure requires sample fixation, slicing, and labeling. Our goal is to apply multiphoton microscopy to efficiently image and quantitatively analyze liver fibrosis specimens bypassing steps required in histological preparation. In this work,...