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Taking Multiphoton Imaging to New Depths

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Dr. Philip G. Smith and Julien Klein, !%Newport Corp.%!

New laser platforms developed to support the latest needs of biological imaging are poised to fuel the next wave of innovation in multiphoton microscopy: imaging deeper into tissue. Two-photon excited fluorescence (TPEF) microscopy has seen rapid growth in the past two decades, fueled by substantial advances in the supporting laser technology. TPEF microscopy was developed for live-cell imaging and uses femtosecond laser pulses for two-photon excitation of fluorophores. This excitation occurs only within a tight focal volume, where the laser’s photon density is high enough to...Read full article

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    Published: October 2011
    Glossary
    autofluorescence
    Autofluorescence refers to the natural emission of fluorescence exhibited by certain biological structures or molecules when exposed to light. Unlike fluorescence that results from the application of external fluorophores or dyes, autofluorescence arises intrinsically from endogenous molecules present in tissues or cells. Key points about autofluorescence: Endogenous emission: Autofluorescence occurs due to the presence of naturally fluorescent molecules within biological samples, such as...
    coherent anti-stokes raman scattering
    A technique whereby two laser beams, one at an excitation wavelength and the second at a wavelength that produces Stokes Raman scattering, interact coherently in a sample, producing a strong scattered beam at the anti-Stokes wavelength.
    second-harmonic generation
    A process whereby two fields of the same optical frequency interact in a nonlinear material to produce a third field, which has a frequency twice that of the two input fields.
    two-photon excited fluorescence
    Two-photon excited fluorescence (TPEF) is a nonlinear optical method that allows imaging of biological cells and living tissue. The advantage of TPEF in comparison to conventional fluorescence microscopy is that it provides natural confocality and allows sectioning of the sample. Because it typically uses near-infrared excitation light, the penetration depth is significantly increased. TPEF is implemented as fast imaging microscopy for noninvasive optical pathology. TPEF has been used in...
    autofluorescenceballistic photonsbiological imagingBiophotonicsCARScoherent anti-Stokes Raman scatteringCornell Universitydispersion compensationdyeselastinFeaturesfemtosecond CARS microscopyfemtosecond lasersfemtosecond Ti:sapphire oscillatorsfluorescent proteinsGFPImagingIn vivo microscopyintrinsic fluorophoresJulien KleinLaser World of Photonicslight scatteringlive-cell imagingMai TaiMicroscopymultimodal imagingmultiphoton imagingmultiphoton microscopyNewport Corp.OPO systemsPhilip G. Smithphoton densitysecond-harmonic generationSensors & DetectorsSpectra-PhysicsTHGthird-harmonic generationthree-photon excited fluorescence microscopyTi:sapphireTPEFTPEF microscopytwo-photon absorptiontwo-photon excited fluorescenceLasers

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