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Combining Second-Harmonic Generation with Multiphoton Imaging

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Angela Goodacre, Dennis Donley and Jiwei Jai, Olympus America Inc.

Second-harmonic generation (SHG) is a label-free method of imaging structures in tissue using a pulsed near-infrared laser. It is generated by the nonabsorptive interaction of two photons with an orderly array of molecules that all point in the same direction. Known as inversion asymmetry, this property of directionality is found in certain types of collagen and elastin fibers, in myosin in muscle myofilaments and in microtubules of the mitotic spindle. Second-harmonic imaging is coherent, meaning that the signal propagates in a fixed direction and is sensitive to excitation light...Read full article

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    Published: October 2010
    Glossary
    fluorescence
    Fluorescence is a type of luminescence, which is the emission of light by a substance that has absorbed light or other electromagnetic radiation. Specifically, fluorescence involves the absorption of light at one wavelength and the subsequent re-emission of light at a longer wavelength. The emitted light occurs almost instantaneously and ceases when the excitation light source is removed. Key characteristics of fluorescence include: Excitation and emission wavelengths: Fluorescent materials...
    photobleaching
    Photobleaching is a phenomenon in which the fluorescence of a fluorophore (a fluorescent molecule or dye) is permanently reduced or eliminated upon prolonged exposure to light. This process occurs due to the photochemical destruction or alteration of the fluorophore molecules, rendering them non-fluorescent. Key points about photobleaching: Mechanism: Photobleaching is typically a result of chemical reactions induced by the absorbed photons. The excessive light exposure causes the...
    rayleigh scattering
    Rayleigh scattering is a phenomenon that occurs when light waves interact with particles or molecules that are much smaller than the wavelength of the light. It is named after the British scientist Lord Rayleigh, who first described the phenomenon in the 19th century. Rayleigh scattering is responsible for the blue color of the sky during the day and the reddish hues seen during sunrise and sunset. The scattering of light occurs because the electric field component of the light waves...
    second-harmonic generation
    Second-harmonic generation (SHG) is a nonlinear optical process that occurs when two photons with the same frequency combine within a nonlinear material, resulting in the generation of a new photon with twice the frequency (and therefore half the wavelength) of the original photons. This phenomenon is a specific case of second-order nonlinear optical effects. Key points about second-harmonic generation include: Nonlinear optical process: SHG is a nonlinear optical effect, meaning that the...
    Angela GoodacrearrayBiophotonicscellularcollagenConsumerDennis Donleydetectorsdipping lensDr. Claudio Vinegonielastin fibersFeaturesFiltersflavinsfluorescencefluorescence imagingfluorescent dyesfluorophoresFluorView FV1000GFPImagingintracellularinversion asymmetrylabel-free methodlaser polarizationlight polarizationliving tissueMassachusetts General HospitalMicroscopymicrotubulesmitotic spindlemolecular orientationmultiphoton excitationmultiphoton imagingmultiphoton systemmusclesmyofilamentmyosinNADHOlympusOlympus America Inc.optical pathphotobleachingphotonsRalph Weissleders Center for Systems BiologyRayleigh scatteringsecond-harmonic generationSensors & DetectorsSHGtissueTPEFtwo-photon excitation fluorescenceYiwei JiaLasers

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