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#Focalpoint cron plus
Since excitation of background fluorescence is avoided, no confocal spatial filter is required, and we obtain all of the advantages of a linear (one-photon) confocal microscope plus the absence of out-of-focus photobleach-ing and photodamage. Two-photon excitation avoids this background in laser-scanning microscopy by virtue of its nonlinear optical absorption character, which almost completely limits the excitation to the high-intensity region at the focal point of the strongly focused excitation laser. Fluorescence microscopy, which can provide submi-cron spatial resolution of chemical dynamics within living cells, is frequently limited in its sensitivity and spatial resolution by background due to out-of-focus fluorescence.
Molecular excitation by two-photon absorption holds great promise for vital imaging of biological systems using laser-scanning microscopy.
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